药学学报

Development name & trade name	Target	Phage display library type and format	Phage display technology	Highest development phase	Indication (s) studied
Adalimumab (D2E7) & Humira®	Tumor necrosis factor (TNF)	Humanization by guided selection, scFv	CAT	FDA approved 2002	Rheumatoid arthritis
Anetumab ravatansine (MF-T/Bay 86-1903, conjugate BAY 94-9343)	Mesothelin	Synthetic, Fab	MorphoSys	Phase Ⅱ	Lung neoplasms, pancreatic cancer, mesothelioma, mesothelin positive
Avelumab	Programmed death-ligand 1 (PD-L1)	Naive, Fab	Dyax	Phase Ⅲ	Triple negative breast neoplasms
BAY1093884	Tissue factor pathway inhibitor (TFPI)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Hemophilia
BAY1129980	C4.4a	Synthetic, scFv	BioInvent	Phase Ⅰ	Advanced solid tumors, neoplasms
BAY1179470	Fibroblast growth factor receptor 2 (FGFR2)	Synthetic, scFv	BioInvent	Phase Ⅰ	Advanced solid tumors, neoplasms
BAY1213790	FXIa	Naive, Fab	Dyax	Phase Ⅰ	Arterial thrombosis; venous thrombosis, knee arthroplasty, total
Belimumab & Benlysta®	B-lymphocyte stimulator (BLyS)	Naive, scFv	CAT	FDA approved 2011	Systemic lupus erythematosus
Bertilimumab (iCo-008/ CT-213)	Eotaxin-1 (CCL-11)	Naive, scFv	CAT	Phase Ⅱ	Pemphigoid, bullous, ulcerative colitis
BHQ880	Dickkopf-1 (DKK1)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Smoldering multiple myeloma
BI-1206 (6G11)	FcgRIIB (CD32B)	Synthetic, scFv	BioInvent	Phase Ⅱ	B-cell lymphoma, chronic lymphocytic leukaemia
BI 836845	Insulin-like growth factor Ⅰ and Ⅱ (IGF-Ⅰ, IGF-Ⅱ)	Naive, Fab	MorphoSys	Phase Ⅰ	Non-small cell lung cancer (NSCLC)
Bimagrumab (BYM338)	Activin A receptor type IIB (ACVR2B)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Sporadic inclusion body myositis
BPS804 (MOR05813)	Sclerostin	Synthetic, Fab	MorphoSys	Phase Ⅱ	Osteoporosis, hypophosphatasia
Briakinumab (ABT874)	Interleukin 12/13	Naive, scFv	CAT	Phase Ⅲ	Moderate to severe plaque psoriasis
Carlumab (CNTO-888)	Chemokine (C-C motif) ligand 2 (CCL2)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Prostate cancer
Cixutumumab (IMCA12)	Insulin-like growth factor-1 receptor (IGF-1R)	Naive, Fab	Dyax	Phase Ⅱ	Different cancer types
Cirutuzumab	Receptor tyrosine kinase like orphan receptor (ROR1)	Naive, Fab		Phase Ⅱ	Chronic lymphocytic leukemia, small lymphocytic lymphoma, mantle-cell lymphoma
CNTO-3175	Toll-like receptor 3 (TLR-3)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Asthma
CNTO-6785	Interleukin 17A (IL17A)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Chronic obstructive pulmonary disease
Darleukin (L-19-IL2)	Extra-domain Bof ﬁbronectin	Semi-synthetic, scfv	‘Pini’ library	Phase Ⅲ	Malignant melanoma
Dekavil (F8-IL10)	Extra-domain A of ﬁbronectin	Semi-synthetic, scfv	ETH-2	Phase Ⅱ	Rheumatoid arthritis
Drozitumab (Apomab, PRO95780)	TRAIL receptor 2 (TRAIL-R2)	ND, scFv	Genentech	Phase Ⅱ	Non-hodgkin's lymphoma, NSCLC
Elgemtumab (LJM716, NOV6)	ERRB3 (HER3)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Head and neck squamous cell carcinoma
Fibromun (L19-TNF)	Anti-EDB human antibody L19 fused to human TNF	Semi-synthetic, scfv	“Pini” library	Phase Ⅲ	Malignant melanoma
Fresolimumab (GC1008)	Transforming growth factor β (TGFβ)	Naive, scfv	CAT	Phase Ⅱ	Primary brain tumors, metastatic breast cancer
Foravirumab (CL184, CR4098)	Rabiesvirus, glycoprotein	Immune, scfv	Crucell	Phase Ⅱ	Rabies
Ganitumab (AMG479)	Insulin-like growth factor receptor (IGF-1R)	Naive, Fab	Dyax	Phase Ⅲ	Metastatic ewing sarcoma
Gantenerumab (R1450)	Amyloid ﬁbrils	Synthetic, Fab	MorphoSys	Phase Ⅲ	Alzheimer's disease
Guselkumab (CNTO1959)	P19 subuntit of interleukin 23	Synthetic, Fab	MorphoSys	Phase Ⅲ	Pustular psoriasis, arthritis, psoriatic
IMC-3C5 (LY3022856)	Vascular endothelial growth factor receptor-3 (VEGFR-3)	Naive, Fab	Dyax	Phase Ⅰ	Neoplasms
Lanadelumab (DX-2930)	Kalikrein	Naive, Fab	Dyax	Phase Ⅲ	Hereditary angioedema
Lexatumumab (HGSETR2)	TRAIL-R2	Naive, scfv	CAT	Phase Ⅰ	Gastroesophageal junction adenocarcinoma
Mapatumumab (HGSETR1)	TNF-related apoptosis, inducing ligand receptor 1 (TRAIL-1)	Naive, scfv	CAT	Phase Ⅱ	Hepatocellular, multiple myeloma, NSCLC
Mavrilimumab (GAM3001)	Granulocytemacrophage-colony stimulating factor receptor (GM-CSF)	Naive, scfv	CAT	Phase Ⅱ	Rheumatoid arthritis
MEDI4212	IgE	Naive, scfv	CAT	Phase Ⅰ	Allergic asthma, atopic dermatitis
MEDI9447	Cluster of differentiation-73 (CD73)	Naive, scfv	CAT	Phase Ⅱ	Carcinoma, NSCLC, ovarian cancer
MOR103 (MOR04357, GSK3196365)	Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Rheumatoid arthritis, multiple sclerosis
MOR202	CD38	Synthetic, Fab	MorphoSys	Phase Ⅱ	Multiple myeloma
Namilumab (MT203)	Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)	Humanization by guided selection, naive (rat CDR3 in human VH), scFv	Micromet	Phase Ⅱ	Plaque psoriasis, rheumatoid arthritis
Necitumumab (IMC11F8) Portrazza®	Epidermal growth factor receptor (EGFR)	Naive, Fab	Dyax	FDA approved 2015	Squamous NSCLC
NI-0501	Interferon-gamma	Naive, scfv	CAT	Phase Ⅲ	Primary haemophagocytic lymphohistiocytosis
Opicinumab (BIIB033)	Leucine rich repeat and immunoglobin-like domain- containing protein 1 (LINGO1)	Naive, Fab	Dyax	Phase Ⅱ	Multiple sclerosis, acute optic neuritis
Oriticumab (BI-204/ MLDL1278A)	Oxidized low-density lipoprotein (oxLDL)	Synthetic, scfv	BioInvent	Phase Ⅱ	Atherosclerosis
Radretumab (L19I, L19SIP)	Extra-domain Bof ﬁbronectin	Semin-synthetic, scfv	“Pini”library	Phase Ⅱ	Brain metastases from solid tumors
Ramucirumab (IMC121B) Cyramaza®	Vascular endothelial growth factor receptor 2 (VEGFR2)	Naive, Fab	Dyax	FDA approved 2014	Gastric cancer, NSCLC
Ranibizumab Lucentis®	Vascular growth factor A (VEGFA)	Afﬁnity maturation of bevacizumab by phage display	Genentech	FDA approved 2006	Macular degeneration
Raxibacumab Abthrax®	Protective (PA)	Naive, scfv	CAT	FDA approved 2012	Anthrax
PC-mAb (M99-B05)	Phosphorylcholine	Naive, Fab	Dyax	Phase Ⅱ	Arterial inflammation, cardiovascular diseases
Seribantumab (MM121)	Epidermal growth factor receptor 3 (ErbB3)	Naive, Fab	Dyax	Phase Ⅱ	Metastatic breast cancer, ER positive, Her2 negative breast cancer patients
Tanibirumab (TTAC0001)	Vascular endothelial growth factor receptor 2 (VEGFR-2)	Naive, scfv	PharmAbcine	Phase Ⅱ	Recurrent glioblastoma
Tarextumab (OMP59R5)	Notch 2/3	Synthetic, Fab	MorphoSys	Phase Ⅱ	Pancreatic cancer, small cell lung cancer
Tleukin (F16-IL2)	Extra-domain A1 of tenascin-C	Semi-synthetic, scfv	ETH-2	Phase Ⅱb	Merkel cell carcinoma, solid tumour, breast cancer
Tesidolumab (LFG316)	Complement C5	Synthetic, Fab	MorphoSys	Phase Ⅱ	Paroxysmal nocturnal hemoglobinuria, geographic atrophy
Tralokinumab (CAT354/BAK1.1)	Interleukin 13	Naive, scFv BAK1 scFv affinity maturation by phage display	CAT	Phase Ⅲ (currently terminated)	Inadequately controlled asthma
Utomilumab (PF05082566)	4-1BB (CD137, TNFRSF9)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Diffuse large B-Cell lymphoma
Vantictumab (OMP18R5)	Frizzled family receptor 7 (Fzd7)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Metastatic breast cancer, solid tumors
VAY-736	B-cell-activating factor receptor (BAFF-R)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Autoimmune hepatitis
Guselkumab Tremfya®	P19 subuntit of interleukin 23 synthetic	MorphoSys, Fab	Morphosys	FDA approved 2017	Pustular psoriasis

Development name & trade name	Target	Phage display library type and format	Phage display technology	Highest development phase	Indication (s) studied
Adalimumab (D2E7) & Humira®	Tumor necrosis factor (TNF)	Humanization by guided selection, scFv	CAT	FDA approved 2002	Rheumatoid arthritis
Anetumab ravatansine (MF-T/Bay 86-1903, conjugate BAY 94-9343)	Mesothelin	Synthetic, Fab	MorphoSys	Phase Ⅱ	Lung neoplasms, pancreatic cancer, mesothelioma, mesothelin positive
Avelumab	Programmed death-ligand 1 (PD-L1)	Naive, Fab	Dyax	Phase Ⅲ	Triple negative breast neoplasms
BAY1093884	Tissue factor pathway inhibitor (TFPI)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Hemophilia
BAY1129980	C4.4a	Synthetic, scFv	BioInvent	Phase Ⅰ	Advanced solid tumors, neoplasms
BAY1179470	Fibroblast growth factor receptor 2 (FGFR2)	Synthetic, scFv	BioInvent	Phase Ⅰ	Advanced solid tumors, neoplasms
BAY1213790	FXIa	Naive, Fab	Dyax	Phase Ⅰ	Arterial thrombosis; venous thrombosis, knee arthroplasty, total
Belimumab & Benlysta®	B-lymphocyte stimulator (BLyS)	Naive, scFv	CAT	FDA approved 2011	Systemic lupus erythematosus
Bertilimumab (iCo-008/ CT-213)	Eotaxin-1 (CCL-11)	Naive, scFv	CAT	Phase Ⅱ	Pemphigoid, bullous, ulcerative colitis
BHQ880	Dickkopf-1 (DKK1)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Smoldering multiple myeloma
BI-1206 (6G11)	FcgRIIB (CD32B)	Synthetic, scFv	BioInvent	Phase Ⅱ	B-cell lymphoma, chronic lymphocytic leukaemia
BI 836845	Insulin-like growth factor Ⅰ and Ⅱ (IGF-Ⅰ, IGF-Ⅱ)	Naive, Fab	MorphoSys	Phase Ⅰ	Non-small cell lung cancer (NSCLC)
Bimagrumab (BYM338)	Activin A receptor type IIB (ACVR2B)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Sporadic inclusion body myositis
BPS804 (MOR05813)	Sclerostin	Synthetic, Fab	MorphoSys	Phase Ⅱ	Osteoporosis, hypophosphatasia
Briakinumab (ABT874)	Interleukin 12/13	Naive, scFv	CAT	Phase Ⅲ	Moderate to severe plaque psoriasis
Carlumab (CNTO-888)	Chemokine (C-C motif) ligand 2 (CCL2)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Prostate cancer
Cixutumumab (IMCA12)	Insulin-like growth factor-1 receptor (IGF-1R)	Naive, Fab	Dyax	Phase Ⅱ	Different cancer types
Cirutuzumab	Receptor tyrosine kinase like orphan receptor (ROR1)	Naive, Fab		Phase Ⅱ	Chronic lymphocytic leukemia, small lymphocytic lymphoma, mantle-cell lymphoma
CNTO-3175	Toll-like receptor 3 (TLR-3)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Asthma
CNTO-6785	Interleukin 17A (IL17A)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Chronic obstructive pulmonary disease
Darleukin (L-19-IL2)	Extra-domain Bof ﬁbronectin	Semi-synthetic, scfv	‘Pini’ library	Phase Ⅲ	Malignant melanoma
Dekavil (F8-IL10)	Extra-domain A of ﬁbronectin	Semi-synthetic, scfv	ETH-2	Phase Ⅱ	Rheumatoid arthritis
Drozitumab (Apomab, PRO95780)	TRAIL receptor 2 (TRAIL-R2)	ND, scFv	Genentech	Phase Ⅱ	Non-hodgkin's lymphoma, NSCLC
Elgemtumab (LJM716, NOV6)	ERRB3 (HER3)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Head and neck squamous cell carcinoma
Fibromun (L19-TNF)	Anti-EDB human antibody L19 fused to human TNF	Semi-synthetic, scfv	“Pini” library	Phase Ⅲ	Malignant melanoma
Fresolimumab (GC1008)	Transforming growth factor β (TGFβ)	Naive, scfv	CAT	Phase Ⅱ	Primary brain tumors, metastatic breast cancer
Foravirumab (CL184, CR4098)	Rabiesvirus, glycoprotein	Immune, scfv	Crucell	Phase Ⅱ	Rabies
Ganitumab (AMG479)	Insulin-like growth factor receptor (IGF-1R)	Naive, Fab	Dyax	Phase Ⅲ	Metastatic ewing sarcoma
Gantenerumab (R1450)	Amyloid ﬁbrils	Synthetic, Fab	MorphoSys	Phase Ⅲ	Alzheimer's disease
Guselkumab (CNTO1959)	P19 subuntit of interleukin 23	Synthetic, Fab	MorphoSys	Phase Ⅲ	Pustular psoriasis, arthritis, psoriatic
IMC-3C5 (LY3022856)	Vascular endothelial growth factor receptor-3 (VEGFR-3)	Naive, Fab	Dyax	Phase Ⅰ	Neoplasms
Lanadelumab (DX-2930)	Kalikrein	Naive, Fab	Dyax	Phase Ⅲ	Hereditary angioedema
Lexatumumab (HGSETR2)	TRAIL-R2	Naive, scfv	CAT	Phase Ⅰ	Gastroesophageal junction adenocarcinoma
Mapatumumab (HGSETR1)	TNF-related apoptosis, inducing ligand receptor 1 (TRAIL-1)	Naive, scfv	CAT	Phase Ⅱ	Hepatocellular, multiple myeloma, NSCLC
Mavrilimumab (GAM3001)	Granulocytemacrophage-colony stimulating factor receptor (GM-CSF)	Naive, scfv	CAT	Phase Ⅱ	Rheumatoid arthritis
MEDI4212	IgE	Naive, scfv	CAT	Phase Ⅰ	Allergic asthma, atopic dermatitis
MEDI9447	Cluster of differentiation-73 (CD73)	Naive, scfv	CAT	Phase Ⅱ	Carcinoma, NSCLC, ovarian cancer
MOR103 (MOR04357, GSK3196365)	Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Rheumatoid arthritis, multiple sclerosis
MOR202	CD38	Synthetic, Fab	MorphoSys	Phase Ⅱ	Multiple myeloma
Namilumab (MT203)	Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)	Humanization by guided selection, naive (rat CDR3 in human VH), scFv	Micromet	Phase Ⅱ	Plaque psoriasis, rheumatoid arthritis
Necitumumab (IMC11F8) Portrazza®	Epidermal growth factor receptor (EGFR)	Naive, Fab	Dyax	FDA approved 2015	Squamous NSCLC
NI-0501	Interferon-gamma	Naive, scfv	CAT	Phase Ⅲ	Primary haemophagocytic lymphohistiocytosis
Opicinumab (BIIB033)	Leucine rich repeat and immunoglobin-like domain- containing protein 1 (LINGO1)	Naive, Fab	Dyax	Phase Ⅱ	Multiple sclerosis, acute optic neuritis
Oriticumab (BI-204/ MLDL1278A)	Oxidized low-density lipoprotein (oxLDL)	Synthetic, scfv	BioInvent	Phase Ⅱ	Atherosclerosis
Radretumab (L19I, L19SIP)	Extra-domain Bof ﬁbronectin	Semin-synthetic, scfv	“Pini”library	Phase Ⅱ	Brain metastases from solid tumors
Ramucirumab (IMC121B) Cyramaza®	Vascular endothelial growth factor receptor 2 (VEGFR2)	Naive, Fab	Dyax	FDA approved 2014	Gastric cancer, NSCLC
Ranibizumab Lucentis®	Vascular growth factor A (VEGFA)	Afﬁnity maturation of bevacizumab by phage display	Genentech	FDA approved 2006	Macular degeneration
Raxibacumab Abthrax®	Protective (PA)	Naive, scfv	CAT	FDA approved 2012	Anthrax
PC-mAb (M99-B05)	Phosphorylcholine	Naive, Fab	Dyax	Phase Ⅱ	Arterial inflammation, cardiovascular diseases
Seribantumab (MM121)	Epidermal growth factor receptor 3 (ErbB3)	Naive, Fab	Dyax	Phase Ⅱ	Metastatic breast cancer, ER positive, Her2 negative breast cancer patients
Tanibirumab (TTAC0001)	Vascular endothelial growth factor receptor 2 (VEGFR-2)	Naive, scfv	PharmAbcine	Phase Ⅱ	Recurrent glioblastoma
Tarextumab (OMP59R5)	Notch 2/3	Synthetic, Fab	MorphoSys	Phase Ⅱ	Pancreatic cancer, small cell lung cancer
Tleukin (F16-IL2)	Extra-domain A1 of tenascin-C	Semi-synthetic, scfv	ETH-2	Phase Ⅱb	Merkel cell carcinoma, solid tumour, breast cancer
Tesidolumab (LFG316)	Complement C5	Synthetic, Fab	MorphoSys	Phase Ⅱ	Paroxysmal nocturnal hemoglobinuria, geographic atrophy
Tralokinumab (CAT354/BAK1.1)	Interleukin 13	Naive, scFv BAK1 scFv affinity maturation by phage display	CAT	Phase Ⅲ (currently terminated)	Inadequately controlled asthma
Utomilumab (PF05082566)	4-1BB (CD137, TNFRSF9)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Diffuse large B-Cell lymphoma
Vantictumab (OMP18R5)	Frizzled family receptor 7 (Fzd7)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Metastatic breast cancer, solid tumors
VAY-736	B-cell-activating factor receptor (BAFF-R)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Autoimmune hepatitis
Guselkumab Tremfya®	P19 subuntit of interleukin 23 synthetic	MorphoSys, Fab	Morphosys	FDA approved 2017	Pustular psoriasis

噬菌体展示技术在药物研发中的应用

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侯雪莹 , 胡卓伟 , 崔冰 ^*

药学学报 | 综述 2018,53(8): 1279-1288

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药学学报 | 综述 2018, 53(8): 1279-1288

噬菌体展示技术在药物研发中的应用

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侯雪莹, 胡卓伟, 崔冰^*

作者信息

中国医学科学院、北京协和医学院药物研究所, 天然药物活性物质与功能国家重点实验室, 北京 100050

通讯作者:

* 崔冰, Tel:86-10-83161187, E-mail:cuibing@imm.ac.cn

Recent advances of phage display techniques for drug discovery

Xue-ying HOU, Zhuo-wei HU, Bing CUI^*

Affiliations

State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China

出版时间: 2018-08-12 doi: 10.16438/j.0513-4870.2018-0527

文章导航

摘要

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噬菌体展示技术利用丝状噬菌体展示蛋白和多肽，从大量变异体中提取所需性质的多肽或蛋白。通过噬菌体展示库技术筛选并获得的抗体片段或多肽，在诊断和治疗中发挥着重要的作用。本文简要介绍噬菌体展示技术原理，主要结合噬菌体展示技术在单抗开发、抗原微生物疫苗研发及多肽类药物中的应用，综述噬菌体展示技术在对肿瘤、自身免疫等多种人类疾病诊断及治疗中的重要作用。

关键词

噬菌体展示 / 单克隆抗体 / 多肽 / 药物研发

Abstract

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Phage display technology utilizes filamentous phage display proteins and polypeptides to extract a desired polypeptide or protein from a large number of variants. The antibody fragments screened and obtained by phage display library technology play an important role in disease diagnosis and treatment. This article briefly introduces the principles of phage display technology, summarizes the development of monoclonal antibodies, the development of antigenic microbial vaccines, and the application of peptide drugs. This review highlights the importance of phage display technology in the diagnosis and treatment of various human diseases such as cancer and autoimmune diseases etc.

Key words

phage display / monoclonal antibody / peptide / drug development

引用本文

侯雪莹, 胡卓伟, 崔冰. 噬菌体展示技术在药物研发中的应用. 药学学报, 2018 , 53 (8) : 1279 -1288 . DOI: 10.16438/j.0513-4870.2018-0527

Xue-ying HOU, Zhuo-wei HU, Bing CUI. Recent advances of phage display techniques for drug discovery[J]. Acta Pharmaceutica Sinica, 2018 , 53 (8) : 1279 -1288 . DOI: 10.16438/j.0513-4870.2018-0527

正文

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噬菌体展示技术(phage display technology)利用基因工程手段将外源基因片段插入噬菌体特定蛋白基因, 通过噬菌体表达外源基因编码的蛋白或多肽, 保持重组融合蛋白的相对空间结构和生物学活性并呈现于噬菌体表面^[1]。构建多样噬菌体库与靶蛋白共同孵育, 经生物淘洗去除非靶蛋白结合型噬菌体株, 通过3～5轮收集、扩增及富集获得高亲和力、高特异性的噬菌体克隆株, 采用基因测序鉴定这些噬菌体克隆编码的蛋白序列可用于进一步研究^[1]。噬菌体展示技术紧密相连表型与基因型, 通过结合重组蛋白识别抗原能力与噬菌体扩增能力形成一项高效表达及筛选体系, 在单克隆抗体研发、肿瘤特异性多肽等生物技术领域展现令人期待的应用前景^{[1, 2]}。本文通过噬菌体展示技术、噬菌体抗体库及多肽库构建及相关技术进展, 综述噬菌体展示技术在单克隆抗体研发、肿瘤特异性多肽筛选及对抗病原微生物疫苗等方面的重要作用。

1 噬菌体展示技术原理

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噬菌体展示系统主要包括丝状噬菌体、T7噬菌体、T4噬菌体与λ噬菌体展示系统, 其中丝状噬菌体展示系统应用最为广泛。非溶源性丝状噬菌体由5个外壳蛋白包裹单链环状DNA组成, 能够在宿主菌中繁殖表达但不破坏宿主菌。Smith^[3]在丝状噬菌体分子生物学研究基础上于20世纪80年代提出噬菌体展示技术。80年代末期, 英美学者同时创建噬菌体抗体库技术。90年代初, McCafferty等^[4]利用噬菌体展示技术成功获得与抗原结合抗体片段, 进一步完善噬菌体抗体库筛选技术。丝状噬菌体具有独特的性状, 主要类型包括M13、f1和fd噬菌体。3种噬菌体核苷酸序列同源性超过98%, 基因组组织形式类似, 同时形成的病毒颗粒大小及性状相近。含有一个单链闭环DNA分子丝状噬菌体基因组由约6 400个核苷酸组成, 基因Ⅷ蛋白是噬菌体颗粒的主要结构蛋白。噬菌体共计约2 700个形态亚单位, 这些亚单位包裹病毒基因组, 形成管状丝状病毒颗粒。丝状噬菌体不在细胞内部组装噬菌体颗粒。基因V蛋白-DNA复合物(正链)移动到细菌细胞膜时, 基因V蛋白从正链脱落, 与出现在被感染细胞膜上的病毒基因组组合形成丝状噬菌体颗粒。被包装的单链DNA在大小上没有严格限制。被噬菌体感染细菌不发生裂解并能够继续缓慢生长, 丝状噬菌体与宿主菌同时进行复制。同一细胞能够产生大量病毒颗粒, 经累积可获得滴度超过1×10¹² pfu·mL^-1噬菌体库。

丝状噬菌体适宜外源重组蛋白表达。F'因子编码的性纤毛在病毒颗粒感染时发挥重要作用, 只有具有性纤毛的菌株才能被病毒颗粒感染。次要衣壳蛋白是基因Ⅲ表达产物, 存在于病毒颗粒丝杆末端。它参与细菌性纤毛末端吸附病毒颗粒过程。丝状噬菌体pⅢ蛋白和pⅧ蛋白处通常用于展示外源性重组蛋白。pⅢ蛋白存在于丝状噬菌体尾端, 外源重组蛋白能够融合在pⅢ蛋白中间部位或N端。如果重组蛋白融合于N端, 则重组噬菌体丧失感染性, 需辅助噬菌体表达完整pⅢ蛋白使其重获感染性。病毒的主要外壳蛋白pⅧ位于丝状噬菌体两侧, 通常N端能够融合五肽。重组更长肽链或蛋白会影响噬菌体装配并导致其丧失感染力。在辅助噬菌体提供pⅧ蛋白前提下, pⅧ蛋白N端能够融合更长的多肽片段甚至抗体片段。此外, 丝状噬菌体pⅣ蛋白的C端亦可作为外源蛋白的融合位点, 多用于外源蛋白C端结构区域功能研究。常用的辅助噬菌体均属于丝状噬菌体, 主要包括含全长基因Ⅲ M13K07、R408、VCSM13等, 及基因Ⅲ删除或缺陷辅助噬菌体等。M13K07是丝状噬菌体M13的突变体, 而VCSM13则是M13K07的突变体。

噬菌粒仅保存噬菌体部分遗传信息, 因此噬菌体库构建及筛选过程中, 宿主细胞的噬菌粒DNA复制和包装中所需的蛋白酶与外壳蛋白均由辅助噬菌体提供^[5]。实际应用过程中, 在辅助噬菌体确保有效感染细菌前提下, 自身需维持低包装效率才能优先包装噬菌体基因组信息。目前, 噬菌体展示技术广泛应用于药物研发的多个领域。本文将着重阐述噬菌体展示技术在单抗开发、多肽类药物发现及抗原微生物疫苗研发等领域的研究进展。

2 噬菌体展示技术在治疗性单克隆抗体开发中的应用

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单克隆抗体(mAbs)在诊断和治疗领域发挥着重要的作用。到目前为止, 欧美已有超过350个治疗性单抗应用于临床研究^[6], 超过50多个抗体已被批准进行销售^[7]。开发具有高亲和力和特异性的新型抗体是目前研究人员面临的挑战。2002年, 阿达木单抗(adalimumab)作为第一个通过噬菌体展示开发的单克隆抗体获得美国食品药品监督管理局批准。此后, 尽管以人源化转基因动物免疫为代表的新抗体开发技术不断涌现, 噬菌体展示技术仍然占据人源化单克隆抗体研发的主导地位^[2]。免疫动物或表面展示是目前单抗开发常用的两种方法。免疫动物接种在抗体研发中曾发挥重要的作用, 但是针对有毒抗原及半抗原的单克隆抗体制备呈现明显缺陷和不足。不同于其他抗体制备手段, 噬菌体抗体展示是一种独立于免疫系统的体外筛选技术。目前常用的技术包括英国剑桥大学Winter研究小组^[8]研发的PHEN1载体构建scFv系统筛选scFv片段, 以及美国Scripps研究所Lerner课题组^[9]研发的pComb3载体构建Fab系统筛选Fab片段。Fab片段及scFv片段均为噬菌体表面展示常见类型, Fab系统可以在大肠杆菌中表达。表 1汇总目前通过噬菌体抗体库开发抗体所使用的展示系统以及抗体适应症等^{[2, 10, 11]}。

噬菌体抗体库是抗体基因经噬菌体展示后形成的多样噬菌体库。噬菌体抗体库的质量主要通过库容量(size)及多样性(diversity)决定。理论上库容达到1×10⁷即可识别超过99%的抗原决定簇。但是, 为获得高亲和力抗体, 噬菌体抗体库在保证多样性前提下还需要尽可能增大库容。噬菌体表面展示的抗体片段数量代表库容大小, 而展示片段的多样性则代表抗体库多样性。噬菌体抗体库库容越大, 筛选获得的抗体亲和力越高。然而, 噬菌粒的转化效率对噬菌体的库容具有重要影响。研究发现, 即使使用库容为1×10¹³的噬菌体库进行建库, 由于较低的细菌转化效率也能造成库容大大降低^[12]。最近的研究通过强力电穿孔、组合感染及Cre-Lox介导重组等方法增加噬菌体库的多样性。

根据噬菌体库的来源, 抗体库主要包括免疫库和合成抗体库。基于抗体基因序列来源, 抗体库又可分为天然库、半合成库和全合成库。合成库亦可根据受体框架、CDR区多样性起源及构建库所用技术进行分类。理论上库容达到1×10⁸的合成抗体库就可筛选到抗原所有可能的抗体片段。天然库是通过未经免疫的动物或人来源的外周血淋巴细胞、骨髓或脾脏细胞, 分离获得B细胞IgM mRNA的V基因。第一个天然库是通过2个健康人外周血淋巴细胞建立^[13]。进一步的研究中, 通过更多捐献者淋巴细胞基因进行组合扩充^{[14, 15]}, 或采用天然库与合成库进行组合扩大库容的同时增加抗体片段的多样性^{[16, 17]}。半合成库不仅包含天然多样性, 同时通过计算机辅助设计针对性增加抗体基因的多样性^[18-20]。Hoogenboom和Winter^[19]通过组合49个不同种系的基因片段制备半合成抗体库。这个scFv抗体库主要通过可变区VH序列和单链Vλ3轻链序列获得, 同时通过CDR-H3区域随机突变5～8个氨基酸进一步增加抗体库的多样性。全合成抗体库通常将种系V基因片段随机插入到CDR区组装, 再通过装配V、D/J基因片段合成构建抗体库^[19]。噬菌体抗体库结合肿瘤细胞体外筛选及体内富集是获得肿瘤特异性抗体的重要模式(图 1)^[21]。最近研究发现, 双特异性抗体(BsAb)能够通过两个结合位点可与同一抗原或两个不同抗原的两个不同抗原表位结合^[22]。不同抗体轻重链之间在单一宿主细胞内存在配对问题。利用这种特性进行BsAb研发, 在噬菌体展示特异性抗人表皮因子受体蛋白(HER2)及血管内皮生长因子(VEGF)抗体研究中, 展示所获抗体不仅保持与原HER2抗原保持高亲和性, 同时识别第二抗原VEGF^{[23, 24]}。

随着噬菌体展示技术广泛应用, 多个单克隆抗体已经应用于临床疾病治疗。这些抗体包括抗肿瘤坏死因子TNF抗体adalimumab (Humira)、抗TNFSF13B抗体belimumab (Benlysta)、抗上皮生长因子受体EGFR抗体necitumumab (Portrazza)、抗血管内皮生长因子VEGF抗体ranibizumab (Lucentis)等^[2]。单克隆抗体噬菌体展示技术专利过期或接近过期(欧洲专利EP0440147、EP0774511、EP0589877;美国专利US5969108、US6172197、US6806079)也将促进更多采用噬菌体展示技术研发的单克隆抗体进入临床。噬菌体展示技术相较于酵母细胞展示或哺乳细胞展示, 不仅有库容大(> 1×10¹⁰)、效力强、表达系统稳定、扩展性强、筛选周期短及费用低等特点, 同时可直接使用抗原表达细胞进行筛选的优势。结合动物免疫库的制备, 噬菌体展示技术可以根据不同的目的灵活进行针对特定抗原的抗体筛选及优化, 或针对不同抗原的单克隆抗体筛选^[2]。

单克隆抗体研发过程中常需要抗体亲和力成熟解决抗体亲和力偏低等类似问题, 噬菌体展示技术常用于体外抗体亲和力成熟。体内抗体亲和力成熟是在体液免疫作用下抗体亲和力逐渐增强的现象, 包括抗体基因重排、体细胞突变及体液免疫3个步骤。抗体基因重排是抗体亲和力成熟的分子基础。B细胞分化过程中其可变区基因在外周发生高频超突变, 克隆选择保存下亲和力高的克隆, 其中一部分转变为记忆细胞。抗体再次刺激记忆细胞时, 重新启动高频突变, 导致高频突变的体细胞数目增多, 同时抗体亲和力增高。

应用噬菌体抗体库展示时, VH与VL基因随机重排模拟抗体体内亲和力成熟的过程, 在此过程中体细胞超突变是限制基础库选择的主要原因。体外亲和力成熟存在多种不同方法, 其中较为常用的方法是增加抗体多样性及增加选择压力^[1]。抗体亲和力增加的方法主要包括非靶向性多样性和靶向性多样性。非靶向多样性增加是通过如mutD5等突变大肠杆菌菌株或错配PCR来实现^{[1, 25, 26]}。实施过程中在抗体骨架的保守区中引入缺失突变, 改变抗体效度, 展示构建更大抗体库进而鉴别高亲和力抗体。PCR错配技术与噬菌体展示技术的结合能够改善潜在抗体亲和性降低及特异性丧失的问题。到目前为止, 利用PCR错配技术和突变大肠杆菌菌株已改良了大量的单克隆抗体。Hawkins等^[27]研究发现, 利用大肠杆菌突变mutD5能够改善特异性抗体phOx的亲和力, 同时使用错配PCR和噬菌体展示技术能够提高抗原特异性抗体的亲和力约4.5倍。非靶向性多样性的另一种常用技术是链改组技术。这种技术将抗体其中一条轻链或重链固定不变用以保持抗体的特异性。同时, 通过将另一条链突变后重新与固定链配对, 反复多次进行链改组和亲和力筛选能够获得更高亲和力的抗体^[28]。研究发现, 对于 > 1 kb DNA基因序列可选用DNA重组结合噬菌体展示技术的方法。定向CDR重组技术是典型靶向多样性增加亲和力的方法。这种技术主要通过在抗体CDR区中引入多种突变或经过设计的DNA序列, 再结合抗体库技术筛选以提高抗体亲和力。然而, 这种方法不能精确解析各CDR区突变作用。在分选高亲和力的FVs、Fabs及重组抗体时, 首选热点突变来提高亲和力。在此基础上, 结合噬菌体库展示技术可诱导抗体亲和力成熟; 利用硅片模型结合噬菌体展示技术, 可避免噬菌体技术库容低的缺点。最近研究发现, 通过CDR诱变技术能够提高抗体对HIV的诊断标志物P24抗原亲和力约5.7倍^[29]。Watanable等^[30]构建非免疫可变区噬菌体展示库, 在目标抗原存在的前提下考察片段复合物稳定性进行筛选。构建抗体VH片段噬菌体展示库, 与可溶性VL片段蛋白及生物素标记的抗原混合后, 稳定三复合物通过streptavidin标记磁珠进行富集及筛选。筛选过程中, 分离与VH突变体具有协同作用的VL突变体, 能够促使抗体亲和性成熟。Sanada等^[31]采用这种方法成功增加抗EGFR抗体528的亲和力。在此基础上, 改进实验方法将可溶性FV片段添置在可被噬菌体库展示的VL片段上。获得新的抗EGFR抗体亲和力与亲本抗体528相当。这种靶向多样性方法能够实现对抗体VL片段的改造及抗体亲和成熟。单克隆抗体在成像和疾病治疗领域逐渐展示出重要作用, 这也促进研究人员通过采用类似噬菌体展示技术等手段研发新型单克隆抗体的重要原因。

3 噬菌体展示技术在多肽类药物研究中的应用

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多肽是在激素、神经递质、细胞因子、抗原和生长因子中参与多种生物过程的生物活性因子^[32]。因此, 在肿瘤学、内分泌学、泌尿学和产科等多个医学领域, 多肽被广泛地应用于治疗和诊断试剂^[33]。目前, 市场上已有超过60种多肽药物被批准, 约140个新型治疗肽处于临床研究^[34]。目前, 已有多个噬菌体展示技术来源多肽获准上市^[35]。美国安进公司获准上市的romiplostim作为血小板生成素受体配基用于治疗特发性血小板减少性紫癜。日本武田公司获准上市的peginesatide作为促红细胞生成素类似物治疗慢性肾病。然而, 多肽存在体内酶促反应快、半衰期短等缺陷。目前主要通过PEG修饰、糖基化和蛋白质结合来改善肽类药物的药代动力学和药效学特性^{[1, 2]}。

多肽噬菌体库展示大量不同蛋白肽模拟真实表位, 这种技术在开发新型治疗多肽领域发挥着重要作用。完全退化的寡核苷酸与噬菌体结构基因融合而成肽噬菌体库。迄今, 采用噬菌体展示技术已经成功研发出多种多肽类药物。例如, 通过噬菌体展示肽库发现的nplate1 (romiplostim, AMG531)是一种肽体(peptibody, 肽-Fc融合蛋白)能够用于治疗免疫血小板减少症。Nplate1是通过4份被识别的肽连接到Fc片段形成融合蛋白延长体内半衰期^[36]。AMG-386 (trebananib)也是通过类似的方式制备的肽体。AMG-386能抑制内皮细胞增殖及肿瘤生长, 目前作为抗癌候选药物进行Ⅲ期临床试验^[37]。此外, 噬菌体展示肽库分离获得的促红细胞生成素受体激动剂CNTO 530能够用于治疗慢性肾病所致贫血。肽段连接人lgG4-Fc片段形成肽体能够获得更长的体内半衰期^[38]。具有酪氨酸激酶活性的HER2在肿瘤发生发展过程中发挥关键作用。Barati等^[39]通过噬菌体展示库获得HER2来源多肽AE37 (li-key/HER-2/neu 776e T60), 同时将AE37与λ噬菌体(λF7)外壳蛋白gpD构建新型纳米颗粒融合肽。融合肽能够产生有效预防和治疗HER2阳性乳腺肿瘤的发展进程。

最近噬菌体展示技术研究发现, 一些多肽类药物具有多种药理生物学活性^[34]。如具有抗菌活性的多功能肽具有可同时促进创面愈合及激活免疫反应的作用^[34]。胰高血糖素类肽-1 (GLP-1)受体激动剂相关多肽药物如albiglutide, 目前作为除运动及饮食控制之外帮助2型糖尿病患者控制血糖的重要辅助手段^[34]。牛流行热病毒引发的牛列热(BEF)能够短时间造成奶牛业的严重经济损失^[40-42]。最近研究发现, 通过病毒特异G1肽蛋白的噬菌体展示肽库T-mer筛选发现抗病毒肽配体T18 (HSIRYDF)及T25 (YSLRSDY)^[43]。纯化BEF病毒G1蛋白后并与M13来源ph-7噬菌体随机显示库共孵育, 针对病毒特异G1肽段进行4轮富集后进行测序。研究者通过ELISA法鉴定阳性克隆形成的多肽, 分析肽配基与BEF病毒特异肽G1的亲和性, 明确病毒感染条件下肽配基仍然保持与G1的结合活力。结果发现, 单独使用T18、T25或联合使用能有效抑制细胞病变斑块的形成, 能够剂量依赖性抑制病毒RNA复制。这些研究为病毒诊断及新型抗病毒药物提供有力的研究工具。

4 噬菌体展示技术在对抗病原微生物疫苗研发的应用

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细菌、真菌、病毒和寄生虫所致传染病仍然是医疗健康保障的潜在威胁。尽管目前已经研制出了多种针对病原微生物的疫苗, 但在全世界范围内传染性疾病仍是导致死亡的主要原因之一。噬菌体展示技术广泛应用于宿主-病原体的相互作用检测、传染疾病标志物诊断、抗病原微生物疫苗及抗体制备等。病原体细胞表面抗原是疫苗研制过程中最合适的靶点, 这些抗原往往直接影响病原菌分裂、复制及毒性^{[44, 45]}。

噬菌体展示技术在感染性疾病领域的应用方法主要包括两种模式:病原靶向噬菌体展示采用细胞复制酶或宿主病原体病毒因子等特定分子靶点进行筛选; 细胞筛选法则选择整个细菌细胞作为筛选靶点^[1]。活病原体的细胞筛选能够靶向天然状态的细胞表面蛋白。这种筛选方法同样适用固相条件下靶抗原不稳定的情况。最近的研究发现, 通过噬菌体展示技术成功开发出多种针对病原微生物的诊断及治疗试剂。这些病原微生物包括幽门螺杆菌^[46]、金葡菌^[47]、铜绿假单胞菌^[48]、利斯马尼亚^[49]和空肠弯曲杆菌^[50]等。

幽门螺杆菌是导致胃炎和消化性溃疡的主要病因。在细菌表面蛋白中, 脲酶作为胃黏膜黏附扩增因子广泛存在所有幽门螺杆菌菌株。这种具有高度免疫原性的抗原是抗体和疫苗开发的理想靶点。研究者构建两个靶向脲酶抑制幽门螺杆菌活性的scFvs, 然后从两个肽库(24肽库和6肽库)筛选能够靶向所构建的scFvs多肽, 这些多肽可以成为潜在的疫苗候选物。筛选结果发现2个肽段能够模拟脲酶抗原表位(TFLPQPRCSALLRYLSEDGVIVPS和YDFYWW), 成为针对幽门螺杆菌的潜在疫苗候选物^{[51, 52]}。

噬菌体展示技术也应用到金葡菌感染的诊断和治疗。金葡菌感染是囊性纤维化儿童呼吸道常见感染^[53]。这种病原菌往往为其他病原体如铜绿假单胞菌提供黏附条件。临床治疗发现, 耐甲氧西林金葡菌(MRSA)的出现增加患者感染发病率和死亡率^{[54, 55]}。最近的研究发现多种控制这种病原体的肽疫苗。RNAⅢ促进金葡菌毒素合成, 而RNAⅢ的生成受RAP (RNAⅢ activating protein)控制。研究者从肽噬菌体库(PhD-12)筛选获得能够模拟RAP的肽段, 成为免疫治疗的潜在候选先导物。肽段免疫小鼠后, 金葡菌再次感染时小鼠可产生针对性的免疫应答^[56]。相同肽库筛选, 研究者发现金葡菌细胞壁肽聚糖(PGN)的模拟肽。该模拟肽能有效引起二次免疫应答或记忆性抗体反应对抗胸腺依赖性的抗原^[57]。此外, 多肽噬菌体库(PhD-12)来源的合成肽(VPHNPGLISLQG)目前也成为诊断金葡菌感染的工具^[58]。

噬菌体展示也成功应用到铜绿假单胞菌疫苗鉴定。铜绿假单胞菌是院内感染的主要原因, 在囊性纤维化、烧伤、器官移植及角膜感染患者中有非常高的致死率^{[59, 60]}。绿脓杆菌耐药菌株传播是目前面临的主要问题。Beckmann等^[61]采用囊性纤维化感染铜绿假单胞菌患者血清, 通过九肽噬菌体文库J404筛选获得多个针对绿脓杆菌具有免疫原性的疫苗候选物。Yuan等^[62]通过多肽噬菌体展示库筛选获得合成多肽Pc-EP能够模拟铜绿假单胞菌的假性膜蛋白HmpA。这段合成肽能够产生IL-1β、IL-6、IL-17和IFN-γ等一系列炎性细胞因子对抗铜绿假单胞菌, 预防铜绿假单胞菌所致角膜炎。

弯曲杆菌(C. jejuni)是鸡传人并引起胃肠道感染的常见原因。控制家禽饲养场的空肠弯曲菌数量能够有效降低人类感染几率。构建对抗弯曲杆菌表面分子的噬菌体展示文库(f88-4/15mer), 筛选获得多个具有抑制空肠弯曲菌功能的多肽先导物^[50]。这些研究工作促进多肽噬菌体展示成功应用人畜共患病的防治。

5 其他

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噬菌体展示技术还可以用于抗体酶(abzymes和catmabs)研发^[1]。抗体酶作为一种催化抗体能够抑制酶转化态或酶活中心。研究发现, 具有选择性蛋白激酶活性的抗体酶能够结合特定蛋白表位, 进而抑制蛋白质相互作用。第一个抗体酶源自哮喘患者血清分离获得的具有蛋白酶活性抗血管活性肠肽(vasoactive intestinal peptide, VIP)抗体酶, 具有迅速水解VIP蛋白的功能。抗体酶与化疗药物结合的抗体导向酶前药疗法(antibody-directed abzyme prodrug therapy, ADAPT)借助噬菌体展示技术, 可以确保抗体一方面结合肿瘤抗原, 另外一方面能够催化释放前药, 进而获得杀伤肿瘤细胞的目的。

6 结语和展望

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噬菌体展示技术作为一种重要的筛选技术在生物医药领域逐渐发挥重要的作用。目前, 通过噬菌体展示技术获得的单克隆抗体及多肽药物正在进行临床前和临床试验。抗体噬菌体展示库获得的初始抗体通常亲和力较低, 通过重组突变增加多样性并经亲和力筛选可获得高亲和力的单克隆抗体。这种策略也是目前获得全人源抗体的常用方法。多肽噬菌体展示库除能够获得疾病模拟肽、酶抑制剂、受体激动剂和拮抗剂外, 逐渐应用于半导体氧化物、稀有金属鉴定和检测等领域。噬菌体展示库的来源、库容、多样性及筛选步骤繁琐度仍是其广泛应用需要关注的问题。噬菌体展示技术在抗体库构建及筛选、抗体亲和力成熟、疫苗及多肽药物研发等领域发挥越来越重要的作用。

基金

收起

中国医学科学院医学与健康科技创新工程(2016-I2M-3-008)
中国医学科学院医学与健康科技创新工程(2016-I2M-1-007)
中国医学科学院中央级公益性科研院所基本科研业务费(2017PT31046)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

Rahbarnia L, Farajnia S, Babaei H, et al. Evolution of phage display technology:from discovery to application[J]. J Drug Target, 2017, 25:216-224.

[2]

Frenzel A, Schirrmann T, Hust M. Phage display-derived human antibodies in clinical development and therapy[J]. MAbs, 2016, 8:1177-1194.

[3]

Smith GP. Filamentous fusion phage:novel expression vectors that display cloned antigens on the virion surface[J]. Science, 1985, 228:1315-1317.

[4]

McCafferty J, Griffiths AD, Winter G, et al. Phage antibodies:filamentous phage displaying antibody variable domains[J]. Nature, 1990, 348:552-554.

[5]

Soltes G, Hust M, Ng KK, et al. On the influence of vector design on antibody phage display[J]. J Biotechnol, 2007, 127:626-637.

[6]

Buss NA, Henderson SJ, McFarlane M, et al. Monoclonal antibody therapeutics:history and future[J]. Curr Opin Pharmacol, 2012, 12:615-622.

[7]

Spiess C, Zhai Q, Carter PJ. Alternative molecular formats and therapeutic applications for bispecific antibodies[J]. Mol Immunol, 2015, 67:95-106.

[8]

Ward ES, Gussow D, Griffiths AD, et al. Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli [J]. Nature, 1989, 341:544- 546.

[9]

Huse WD, Sastry L, Iverson SA, et al. Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda[J]. Science, 1989, 246:1275-1281.

[10]

Choi MY, Widhopf GF 2nd, Wu CC, et al. Pre-clinical specificity and safety of UC-961, a first-in-class monoclonal antibody targeting ROR1[J]. Clin Lymphoma Myeloma Leuk, 2015, 15 Suppl:S167-S169.

[11]

Cui B, Widhopf GF, Prussak CE, et al. Cirmtuzumab vedotin (UC-961ADC3), an anti-ROR1-monomethyl auristatin E antibody-drug conjugate, is a potential treatment for ROR1- positive leukemia and solid tumors[J]. Blood, 2013, 122:1637-1637.

[12]

Sun W, Lin H, Hua F, et al. Optimizing the host bacteria to make a large naive phage antibody library in the recombination system[J]. Acta Pharm Sin (药学学报), 2013, 48:66-70.

[13]

Marks JD, Hoogenboom HR, Bonnert TP, et al. By-passing immunization. human antibodies from V-gene libraries displayed on phage[J]. J Mol Biol, 1991, 222:581-597.

[14]

Vaughan TJ, Williams AJ, Pritchard K, et al. Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library[J]. Nat Biotechnol, 1996, 14:309-314.

[15]

de Haard HJ, van Neer N, Reurs A, et al. A large non- immunized human Fab fragment phage library that permits rapid isolation and kinetic analysis of high affinity antibodies[J]. J Biol Chem, 1999, 274:18218-18230.

[16]

Griffiths AD, Williams SC, Hartley O, et al. Isolation of high affinity human antibodies directly from large synthetic repertoires[J]. EMBO J, 1994, 13:3245-3260.

[17]

Hoet RM, Cohen EH, Kent RB, et al. Generation of high- affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity[J]. Nat Biotechnol, 2005, 23:344-348.

[18]

Pini A, Viti F, Santucci A, et al. Design and use of a phage display library. Human antibodies with subnanomolar affinity against a marker of angiogenesis eluted from a two-dimensional gel[J]. J Biol Chem, 1998, 273:21769-21776.

[19]

Hoogenboom HR, Winter G. By-passing immunisation. Human antibodies from synthetic repertoires of germline VH gene segments rearranged in vitro[J]. J Mol Biol, 1992, 227:381-388.

[20]

Nissim A, Hoogenboom HR, Tomlinson IM, et al. Antibody fragments from a 'single pot' phage display library as immunochemical reagents[J]. EMBO J, 1994, 13:692-698.

[21]

Sanchez-Martin D, Sorensen MD, Lykkemark S, et al. Selec tion strategies for anticancer antibody discovery:searching off the beaten path[J]. Trends Biotechnol, 2015, 33:292-301.

[22]

Byrne H, Conroy PJ, Whisstock JC, et al. A tale of two specificities:bispecific antibodies for therapeutic and diagnostic applications[J]. Trends Biotechnol, 2013, 31:621-632.

[23]

Valladares IG, Espinoza LR. Designing two-in-one antibodies[J]. Immunotherapy, 2009, 1:749-751.

[24]

Kontermann RE, Brinkmann U. Bispecific antibodies[J]. Drug Discov Today, 2015, 20:838-847.

[25]

Kilcher S, Studer P, Muessner C, et al. Cross-genus rebooting of custom-made, synthetic bacteriophage genomes in L-form bacteria[J]. Proc Nat Acad Sci U S A, 2018, 115:567-572.

[26]

Unkauf T, Hust M, Frenzel A. Antibody affinity and stability maturation by error-prone PCR[J]. Methods Mol Biol, 2018, 1701:393-407.

[27]

Hawkins RE, Russell SJ, Winter G. Selection of phage anti bodies by binding affinity:mimicking affinity maturation[J]. J Mol Biol, 1992, 226:889-896.

[28]

Stemmer WP. Rapid evolution of a protein in vitro by DNA shuffling[J]. Nature, 1994, 370:389-391.

[29]

Xia L, Zhang J, Cui C, et al. In vitro affinity maturation and characterization of anti-P24 antibody for HIV diagnostic assay[J]. J Biochem, 2015, 158:531-538.

[30]

Watanabe H, Tsumoto K, Asano R, et al. Selection of human antibody fragments on the basis of stabilization of the variable domain in the presence of target antigens[J]. Biochem Biophys Res Commun, 2002, 295:31-36.

[31]

Sanada H, Kobayashi K, Oyama K, et al. Affinity maturation of humanized anti-epidermal growth factor receptor antibody using a modified phage-based open sandwich selection method[J]. Sci Rep, 2018, 8:5414.

[32]

Morishita M, Peppas NA. Is the oral route possible for peptide and protein drug delivery?[J]. Drug Discov Today, 2006, 11:905-910.

[33]

Vergote V, Burvenich C, Van de Wiele C, et al. Quality specifications for peptide drugs:a regulatory-pharmaceutical approach[J]. J Pept Sci, 2009, 15:697-710.

[34]

Fosgerau K, Hoffmann T. Peptide therapeutics:current status and future directions[J]. Drug Discov Today, 2015, 20:122-128.

[35]

Omidfar K, Daneshpour M. Advances in phage display technology for drug discovery[J]. Exp Opin Drug Discov, 2015, 10:651-669.

[36]

Bussel JB, Kuter DJ, George JN, et al. AMG 531, a thrombopoiesis-stimulating protein, for chronic ITP[J]. New Engl J Med, 2006, 355:1672-1681.

[37]

Moghaddam A, Borgen T, Stacy J, et al. Identification of scFv antibody fragments that specifically recognise the heroin metabolite 6-monoacetylmorphine but not morphine[J]. J Immunol Methods, 2003, 280:139-155.

[38]

Wu B, Sun YN. Pharmacokinetics of peptide-Fc fusion proteins[J]. J Pharm Sci, 2014, 103:53-64.

[39]

Barati N, Razazan A, Nicastro J, et al. Immunogenicity and antitumor activity of the superlytic λF7 phage nanoparticles displaying a HER2/neu-derived peptide AE37 in a tumor model of BALB/c mice[J]. Cancer Lett, 2018, 424:109- 116.

[40]

Uren MF. Bovine ephemeral fever[J]. Aust Vet J, 1989, 66:233-236.

[41]

Wang FI, Hsu AM, Huang KJ. Bovine ephemeral fever in Taiwan[J]. J Vet Diagn Invest, 2001, 13:462-467.

[42]

Walker PJ. Bovine ephemeral fever in Australia and the world[J]. Curr Top Microbiol Immunol, 2005, 292:57-80.

[43]

Hou P, Zhao G, He C, et al. Biopanning of polypeptides binding to bovine ephemeral fever virus G1 protein from phage display peptide library[J]. BMC Vet Res, 2018, 14:3.

[44]

Lock RL, Harry EJ. Cell-division inhibitors:new insights for future antibiotics[J]. Nat Rev Drug Discov, 2008, 7:324-338.

[45]

Rasko DA, Sperandio V. Anti-virulence strategies to combat bacteria-mediated disease[J]. Nat Rev Drug Discov, 2010, 9:117-128.

[46]

Ardekani LS, Gargari SL, Rasooli I, et al. A novel nanobody against urease activity of Helicobacter pylori[J]. Int J Infect Dis, 2013, 17:e723-e728.

[47]

O'Rourke JP, Daly SM, Triplett KD, et al. Development of a mimotope vaccine targeting the Staphylococcus aureus quorum sensing pathway[J]. PLoS One, 2014, 9:e111198.

[48]

Secor PR, Michaels LA, Smigiel KS, et al. Filamentous bacteriophage produced by Pseudomonas aeruginosa alters the inflammatory response and promotes noninvasive infection in vivo [J]. Infect Immun, 2017, 85:e00648-16.

[49]

Coelho EA, Chavez-Fumagalli MA, Costa LE, et al. Ther anostic applications of phage display to control leishmaniasis:selection of biomarkers for serodiagnostics, vaccination, and immunotherapy[J]. Rev Soc Bras Med Trop, 2015, 48:370- 379.

[50]

Bishop-Hurley SL, Rea PJ, McSweeney CS. Phage-displayed peptides selected for binding to Campylobacter jejuni are antimicrobial[J]. Protein Eng Des Sel, 2010, 23:751-757.

[51]

Houimel M, Mach JP, Corthesy-Theulaz I, et al. New inhibitors of Helicobacter pylori urease holoenzyme selected from phage-displayed peptide libraries[J]. Eur J Biochem, 1999, 262:774-780.

[52]

Keller WC, Michetti P. Vaccination against Helicobacter pylori——an old companion of man[J]. Expert Opin Biol Ther, 2001, 1:795-802.

[53]

Ahlgren HG, Benedetti A, Landry JS, et al. Clinical outcomes associated with Staphylococcus aureus and Pseudomonas aeruginosa airway infections in adult cystic fibrosis patients[J]. BMC Pulm Med, 2015, 15:67.

[54]

Appelbaum PC. Microbiology of antibiotic resistance in Staphylococcus aureus[J]. Clin Infect Dis, 2007, 45 Suppl 3:S165-S170.

[55]

Aswani V, Mau B, Shukla SK. Complete genome sequence of Staphylococcus aureus MCRF184, a necrotizing fasciitis- causing methicillin-sensitive sequence type 45Staphylococcus strain[J]. Genome Announc, 2016. DOI:10.1128/genome A.00374-16.

[56]

Yang G, Gao Y, Dong J, et al. A novel peptide isolated from phage library to substitute a complex system for a vaccine against staphylococci infection[J]. Vaccine, 2006, 24:1117- 1123.

[57]

Chen Y, Liu B, Yang D, et al. Peptide mimics of peptidoglycan are vaccine candidates and protect mice from infection with Staphylococcus aureus[J]. J Med Microbiol, 2011, 60:995- 1002.

[58]

Rao SS, Mohan KV, Gao Y, et al. Identification and evaluation of a novel peptide binding to the cell surface of Staphylococcus aureus[J]. Microbiol Res, 2013, 168:106-112.

[59]

Crousilles A, Maunders E, Bartlett S, et al. Which microbial factors really are important in Pseudomonas aeruginosa infections?[J]. Future Microbiol, 2015, 10:1825-1836.

[60]

Hakki M, Limaye AP, Kim HW, et al. Invasive Pseudomonas aeruginosa infections:high rate of recurrence and mortality after hematopoietic cell transplantation[J]. Bone Marrow Transplant, 2007, 39:687-693.

[61]

Beckmann C, Brittnacher M, Ernst R, et al. Use of phage display to identify potential Pseudomonas aeruginosa gene products relevant to early cystic fibrosis airway infections[J]. Infect Immun, 2005, 73:444-452.

[62]

Yuan Q, Wu Y, Wang Y, et al. Protective efficacy of a peptide derived from a potential adhesin of Pseudomonas aeruginosa against corneal infection[J]. Exp Eye Res, 2016, 143:39- 48.

2018年第53卷第8期

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doi: 10.16438/j.0513-4870.2018-0527

接收时间：2018-06-05
首发时间：2026-01-15
出版时间：2018-08-12

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收稿日期：2018-06-05
修回日期：2018-06-22

基金

中国医学科学院医学与健康科技创新工程(2016-I2M-3-008)

中国医学科学院医学与健康科技创新工程(2016-I2M-1-007)

中国医学科学院中央级公益性科研院所基本科研业务费(2017PT31046)

作者信息

中国医学科学院、北京协和医学院药物研究所, 天然药物活性物质与功能国家重点实验室, 北京 100050

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* 崔冰, Tel:86-10-83161187, E-mail:cuibing@imm.ac.cn

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Development name & trade name	Target	Phage display library type and format	Phage display technology	Highest development phase	Indication (s) studied
Adalimumab (D2E7) & Humira®	Tumor necrosis factor (TNF)	Humanization by guided selection, scFv	CAT	FDA approved 2002	Rheumatoid arthritis
Anetumab ravatansine (MF-T/Bay 86-1903, conjugate BAY 94-9343)	Mesothelin	Synthetic, Fab	MorphoSys	Phase Ⅱ	Lung neoplasms, pancreatic cancer, mesothelioma, mesothelin positive
Avelumab	Programmed death-ligand 1 (PD-L1)	Naive, Fab	Dyax	Phase Ⅲ	Triple negative breast neoplasms
BAY1093884	Tissue factor pathway inhibitor (TFPI)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Hemophilia
BAY1129980	C4.4a	Synthetic, scFv	BioInvent	Phase Ⅰ	Advanced solid tumors, neoplasms
BAY1179470	Fibroblast growth factor receptor 2 (FGFR2)	Synthetic, scFv	BioInvent	Phase Ⅰ	Advanced solid tumors, neoplasms
BAY1213790	FXIa	Naive, Fab	Dyax	Phase Ⅰ	Arterial thrombosis; venous thrombosis, knee arthroplasty, total
Belimumab & Benlysta®	B-lymphocyte stimulator (BLyS)	Naive, scFv	CAT	FDA approved 2011	Systemic lupus erythematosus
Bertilimumab (iCo-008/ CT-213)	Eotaxin-1 (CCL-11)	Naive, scFv	CAT	Phase Ⅱ	Pemphigoid, bullous, ulcerative colitis
BHQ880	Dickkopf-1 (DKK1)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Smoldering multiple myeloma
BI-1206 (6G11)	FcgRIIB (CD32B)	Synthetic, scFv	BioInvent	Phase Ⅱ	B-cell lymphoma, chronic lymphocytic leukaemia
BI 836845	Insulin-like growth factor Ⅰ and Ⅱ (IGF-Ⅰ, IGF-Ⅱ)	Naive, Fab	MorphoSys	Phase Ⅰ	Non-small cell lung cancer (NSCLC)
Bimagrumab (BYM338)	Activin A receptor type IIB (ACVR2B)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Sporadic inclusion body myositis
BPS804 (MOR05813)	Sclerostin	Synthetic, Fab	MorphoSys	Phase Ⅱ	Osteoporosis, hypophosphatasia
Briakinumab (ABT874)	Interleukin 12/13	Naive, scFv	CAT	Phase Ⅲ	Moderate to severe plaque psoriasis
Carlumab (CNTO-888)	Chemokine (C-C motif) ligand 2 (CCL2)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Prostate cancer
Cixutumumab (IMCA12)	Insulin-like growth factor-1 receptor (IGF-1R)	Naive, Fab	Dyax	Phase Ⅱ	Different cancer types
Cirutuzumab	Receptor tyrosine kinase like orphan receptor (ROR1)	Naive, Fab		Phase Ⅱ	Chronic lymphocytic leukemia, small lymphocytic lymphoma, mantle-cell lymphoma
CNTO-3175	Toll-like receptor 3 (TLR-3)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Asthma
CNTO-6785	Interleukin 17A (IL17A)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Chronic obstructive pulmonary disease
Darleukin (L-19-IL2)	Extra-domain Bof ﬁbronectin	Semi-synthetic, scfv	‘Pini’ library	Phase Ⅲ	Malignant melanoma
Dekavil (F8-IL10)	Extra-domain A of ﬁbronectin	Semi-synthetic, scfv	ETH-2	Phase Ⅱ	Rheumatoid arthritis
Drozitumab (Apomab, PRO95780)	TRAIL receptor 2 (TRAIL-R2)	ND, scFv	Genentech	Phase Ⅱ	Non-hodgkin's lymphoma, NSCLC
Elgemtumab (LJM716, NOV6)	ERRB3 (HER3)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Head and neck squamous cell carcinoma
Fibromun (L19-TNF)	Anti-EDB human antibody L19 fused to human TNF	Semi-synthetic, scfv	“Pini” library	Phase Ⅲ	Malignant melanoma
Fresolimumab (GC1008)	Transforming growth factor β (TGFβ)	Naive, scfv	CAT	Phase Ⅱ	Primary brain tumors, metastatic breast cancer
Foravirumab (CL184, CR4098)	Rabiesvirus, glycoprotein	Immune, scfv	Crucell	Phase Ⅱ	Rabies
Ganitumab (AMG479)	Insulin-like growth factor receptor (IGF-1R)	Naive, Fab	Dyax	Phase Ⅲ	Metastatic ewing sarcoma
Gantenerumab (R1450)	Amyloid ﬁbrils	Synthetic, Fab	MorphoSys	Phase Ⅲ	Alzheimer's disease
Guselkumab (CNTO1959)	P19 subuntit of interleukin 23	Synthetic, Fab	MorphoSys	Phase Ⅲ	Pustular psoriasis, arthritis, psoriatic
IMC-3C5 (LY3022856)	Vascular endothelial growth factor receptor-3 (VEGFR-3)	Naive, Fab	Dyax	Phase Ⅰ	Neoplasms
Lanadelumab (DX-2930)	Kalikrein	Naive, Fab	Dyax	Phase Ⅲ	Hereditary angioedema
Lexatumumab (HGSETR2)	TRAIL-R2	Naive, scfv	CAT	Phase Ⅰ	Gastroesophageal junction adenocarcinoma
Mapatumumab (HGSETR1)	TNF-related apoptosis, inducing ligand receptor 1 (TRAIL-1)	Naive, scfv	CAT	Phase Ⅱ	Hepatocellular, multiple myeloma, NSCLC
Mavrilimumab (GAM3001)	Granulocytemacrophage-colony stimulating factor receptor (GM-CSF)	Naive, scfv	CAT	Phase Ⅱ	Rheumatoid arthritis
MEDI4212	IgE	Naive, scfv	CAT	Phase Ⅰ	Allergic asthma, atopic dermatitis
MEDI9447	Cluster of differentiation-73 (CD73)	Naive, scfv	CAT	Phase Ⅱ	Carcinoma, NSCLC, ovarian cancer
MOR103 (MOR04357, GSK3196365)	Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)	Synthetic, Fab	MorphoSys	Phase Ⅱ	Rheumatoid arthritis, multiple sclerosis
MOR202	CD38	Synthetic, Fab	MorphoSys	Phase Ⅱ	Multiple myeloma
Namilumab (MT203)	Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)	Humanization by guided selection, naive (rat CDR3 in human VH), scFv	Micromet	Phase Ⅱ	Plaque psoriasis, rheumatoid arthritis
Necitumumab (IMC11F8) Portrazza®	Epidermal growth factor receptor (EGFR)	Naive, Fab	Dyax	FDA approved 2015	Squamous NSCLC
NI-0501	Interferon-gamma	Naive, scfv	CAT	Phase Ⅲ	Primary haemophagocytic lymphohistiocytosis
Opicinumab (BIIB033)	Leucine rich repeat and immunoglobin-like domain- containing protein 1 (LINGO1)	Naive, Fab	Dyax	Phase Ⅱ	Multiple sclerosis, acute optic neuritis
Oriticumab (BI-204/ MLDL1278A)	Oxidized low-density lipoprotein (oxLDL)	Synthetic, scfv	BioInvent	Phase Ⅱ	Atherosclerosis
Radretumab (L19I, L19SIP)	Extra-domain Bof ﬁbronectin	Semin-synthetic, scfv	“Pini”library	Phase Ⅱ	Brain metastases from solid tumors
Ramucirumab (IMC121B) Cyramaza®	Vascular endothelial growth factor receptor 2 (VEGFR2)	Naive, Fab	Dyax	FDA approved 2014	Gastric cancer, NSCLC
Ranibizumab Lucentis®	Vascular growth factor A (VEGFA)	Afﬁnity maturation of bevacizumab by phage display	Genentech	FDA approved 2006	Macular degeneration
Raxibacumab Abthrax®	Protective (PA)	Naive, scfv	CAT	FDA approved 2012	Anthrax
PC-mAb (M99-B05)	Phosphorylcholine	Naive, Fab	Dyax	Phase Ⅱ	Arterial inflammation, cardiovascular diseases
Seribantumab (MM121)	Epidermal growth factor receptor 3 (ErbB3)	Naive, Fab	Dyax	Phase Ⅱ	Metastatic breast cancer, ER positive, Her2 negative breast cancer patients
Tanibirumab (TTAC0001)	Vascular endothelial growth factor receptor 2 (VEGFR-2)	Naive, scfv	PharmAbcine	Phase Ⅱ	Recurrent glioblastoma
Tarextumab (OMP59R5)	Notch 2/3	Synthetic, Fab	MorphoSys	Phase Ⅱ	Pancreatic cancer, small cell lung cancer
Tleukin (F16-IL2)	Extra-domain A1 of tenascin-C	Semi-synthetic, scfv	ETH-2	Phase Ⅱb	Merkel cell carcinoma, solid tumour, breast cancer
Tesidolumab (LFG316)	Complement C5	Synthetic, Fab	MorphoSys	Phase Ⅱ	Paroxysmal nocturnal hemoglobinuria, geographic atrophy
Tralokinumab (CAT354/BAK1.1)	Interleukin 13	Naive, scFv BAK1 scFv affinity maturation by phage display	CAT	Phase Ⅲ (currently terminated)	Inadequately controlled asthma
Utomilumab (PF05082566)	4-1BB (CD137, TNFRSF9)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Diffuse large B-Cell lymphoma
Vantictumab (OMP18R5)	Frizzled family receptor 7 (Fzd7)	Synthetic, Fab	MorphoSys	Phase Ⅰ	Metastatic breast cancer, solid tumors
VAY-736	B-cell-activating factor receptor (BAFF-R)	Synthetic, Fab	MorphoSys	Phase Ⅲ	Autoimmune hepatitis
Guselkumab Tremfya®	P19 subuntit of interleukin 23 synthetic	MorphoSys, Fab	Morphosys	FDA approved 2017	Pustular psoriasis

Development name & trade name

Target

Phage display library type and format

Phage display technology

Highest development phase

Indication (s) studied

Adalimumab (D2E7) & Humira®

Tumor necrosis factor (TNF)

Humanization by guided selection, scFv

CAT

FDA approved 2002

Rheumatoid arthritis

Anetumab ravatansine (MF-T/Bay 86-1903, conjugate BAY 94-9343)

Mesothelin

Synthetic, Fab

MorphoSys

Phase Ⅱ

Lung neoplasms, pancreatic cancer, mesothelioma, mesothelin positive

Avelumab

Programmed death-ligand 1 (PD-L1)

Naive, Fab

Dyax

Phase Ⅲ

Triple negative breast neoplasms

BAY1093884

Tissue factor pathway inhibitor (TFPI)

Synthetic, Fab

MorphoSys

Phase Ⅰ

Hemophilia

BAY1129980

C4.4a

Synthetic, scFv

BioInvent

Phase Ⅰ

Advanced solid tumors, neoplasms

BAY1179470

Fibroblast growth factor receptor 2 (FGFR2)

Synthetic, scFv

BioInvent

Phase Ⅰ

Advanced solid tumors, neoplasms

BAY1213790

FXIa

Naive, Fab

Dyax

Phase Ⅰ

Arterial thrombosis; venous thrombosis, knee arthroplasty, total

Belimumab & Benlysta®

B-lymphocyte stimulator (BLyS)

Naive, scFv

CAT

FDA approved 2011

Systemic lupus erythematosus

Bertilimumab (iCo-008/ CT-213)

Eotaxin-1 (CCL-11)

Naive, scFv

CAT

Phase Ⅱ

Pemphigoid, bullous, ulcerative colitis

BHQ880

Dickkopf-1 (DKK1)

Synthetic, Fab

MorphoSys

Phase Ⅱ

Smoldering multiple myeloma

BI-1206 (6G11)

FcgRIIB (CD32B)

Synthetic, scFv

BioInvent

Phase Ⅱ

B-cell lymphoma, chronic lymphocytic leukaemia

BI 836845

Insulin-like growth factor Ⅰ and Ⅱ (IGF-Ⅰ, IGF-Ⅱ)

Naive, Fab

MorphoSys

Phase Ⅰ

Non-small cell lung cancer (NSCLC)

Bimagrumab (BYM338)

Activin A receptor type IIB (ACVR2B)

Synthetic, Fab

MorphoSys

Phase Ⅲ

Sporadic inclusion body myositis

BPS804 (MOR05813)

Sclerostin

Synthetic, Fab

MorphoSys

Phase Ⅱ

Osteoporosis, hypophosphatasia

Briakinumab (ABT874)

Interleukin 12/13

Naive, scFv

CAT

Phase Ⅲ

Moderate to severe plaque psoriasis

Carlumab (CNTO-888)

Chemokine (C-C motif) ligand 2 (CCL2)

Synthetic, Fab

MorphoSys

Phase Ⅱ

Prostate cancer

Cixutumumab (IMCA12)

Insulin-like growth factor-1 receptor (IGF-1R)

Naive, Fab

Dyax

Phase Ⅱ

Different cancer types

Cirutuzumab

Receptor tyrosine kinase like orphan receptor (ROR1)

Naive, Fab

Phase Ⅱ

Chronic lymphocytic leukemia, small lymphocytic lymphoma, mantle-cell lymphoma

CNTO-3175

Toll-like receptor 3 (TLR-3)

Synthetic, Fab

MorphoSys

Phase Ⅰ

Asthma

CNTO-6785

Interleukin 17A (IL17A)

Synthetic, Fab

MorphoSys

Phase Ⅱ

Chronic obstructive pulmonary disease

Darleukin (L-19-IL2)

Extra-domain Bof ﬁbronectin

Semi-synthetic, scfv

‘Pini’ library

Phase Ⅲ

Malignant melanoma

Dekavil (F8-IL10)

Extra-domain A of ﬁbronectin

Semi-synthetic, scfv

ETH-2

Phase Ⅱ

Rheumatoid arthritis

Drozitumab (Apomab, PRO95780)

TRAIL receptor 2 (TRAIL-R2)

ND, scFv

Genentech

Phase Ⅱ

Non-hodgkin's lymphoma, NSCLC

Elgemtumab (LJM716, NOV6)

ERRB3 (HER3)

Synthetic, Fab

MorphoSys

Phase Ⅱ

Head and neck squamous cell carcinoma

Fibromun (L19-TNF)

Anti-EDB human antibody L19 fused to human TNF

Semi-synthetic, scfv

“Pini” library

Phase Ⅲ

Malignant melanoma

Fresolimumab (GC1008)

Transforming growth factor β (TGFβ)

Naive, scfv

CAT

Phase Ⅱ

Primary brain tumors, metastatic breast cancer

Foravirumab (CL184, CR4098)

Rabiesvirus, glycoprotein

Immune, scfv

Crucell

Phase Ⅱ

Rabies

Ganitumab (AMG479)

Insulin-like growth factor receptor (IGF-1R)

Naive, Fab

Dyax

Phase Ⅲ

Metastatic ewing sarcoma

Gantenerumab (R1450)

Amyloid ﬁbrils

Synthetic, Fab

MorphoSys

Phase Ⅲ

Alzheimer's disease

Guselkumab (CNTO1959)

P19 subuntit of interleukin 23

Synthetic, Fab

MorphoSys

Phase Ⅲ

Pustular psoriasis, arthritis, psoriatic

IMC-3C5 (LY3022856)

Vascular endothelial growth factor receptor-3 (VEGFR-3)

Naive, Fab

Dyax

Phase Ⅰ

Neoplasms

Lanadelumab (DX-2930)

Kalikrein

Naive, Fab

Dyax

Phase Ⅲ

Hereditary angioedema

Lexatumumab (HGSETR2)

TRAIL-R2

Naive, scfv

CAT

Phase Ⅰ

Gastroesophageal junction adenocarcinoma

Mapatumumab (HGSETR1)

TNF-related apoptosis, inducing ligand receptor 1 (TRAIL-1)

Naive, scfv

CAT

Phase Ⅱ

Hepatocellular, multiple myeloma, NSCLC

Mavrilimumab (GAM3001)

Granulocytemacrophage-colony stimulating factor receptor (GM-CSF)

Naive, scfv

CAT

Phase Ⅱ

Rheumatoid arthritis

MEDI4212

IgE

Naive, scfv

CAT

Phase Ⅰ

Allergic asthma, atopic dermatitis

MEDI9447

Cluster of differentiation-73 (CD73)

Naive, scfv

CAT

Phase Ⅱ

Carcinoma, NSCLC, ovarian cancer

MOR103 (MOR04357, GSK3196365)

Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)

Synthetic, Fab

MorphoSys

Phase Ⅱ

Rheumatoid arthritis, multiple sclerosis

MOR202

CD38

Synthetic, Fab

MorphoSys

Phase Ⅱ

Multiple myeloma

Namilumab (MT203)

Granulocyte macrophage-colony stimulating factor receptor (GM-CSF)

Humanization by guided selection, naive (rat CDR3 in human VH), scFv

Micromet

Phase Ⅱ

Plaque psoriasis, rheumatoid arthritis

Necitumumab (IMC11F8) Portrazza®

Epidermal growth factor receptor (EGFR)

Naive, Fab

Dyax

FDA approved 2015

Squamous NSCLC

NI-0501

Interferon-gamma

Naive, scfv

CAT

Phase Ⅲ

Primary haemophagocytic lymphohistiocytosis

Opicinumab (BIIB033)

Leucine rich repeat and immunoglobin-like domain- containing protein 1 (LINGO1)

Naive, Fab

Dyax

Phase Ⅱ

Multiple sclerosis, acute optic neuritis

Oriticumab (BI-204/ MLDL1278A)

Oxidized low-density lipoprotein (oxLDL)

Synthetic, scfv

BioInvent

Phase Ⅱ

Atherosclerosis

Radretumab (L19I, L19SIP)

Extra-domain Bof ﬁbronectin

Semin-synthetic, scfv

“Pini”library

Phase Ⅱ

Brain metastases from solid tumors

Ramucirumab (IMC121B) Cyramaza®

Vascular endothelial growth factor receptor 2 (VEGFR2)

Naive, Fab

Dyax

FDA approved 2014

Gastric cancer, NSCLC

Ranibizumab Lucentis®

Vascular growth factor A (VEGFA)

Afﬁnity maturation of bevacizumab by phage display

Genentech

FDA approved 2006

Macular degeneration

Raxibacumab Abthrax®

Protective (PA)

Naive, scfv

CAT

FDA approved 2012

Anthrax

PC-mAb (M99-B05)

Phosphorylcholine

Naive, Fab

Dyax

Phase Ⅱ

Arterial inflammation, cardiovascular diseases

Seribantumab (MM121)

Epidermal growth factor receptor 3 (ErbB3)

Naive, Fab

Dyax

Phase Ⅱ

Metastatic breast cancer, ER positive, Her2 negative breast cancer patients

Tanibirumab (TTAC0001)

Vascular endothelial growth factor receptor 2 (VEGFR-2)

Naive, scfv

PharmAbcine

Phase Ⅱ

Recurrent glioblastoma

Tarextumab (OMP59R5)

Notch 2/3

Synthetic, Fab

MorphoSys

Phase Ⅱ

Pancreatic cancer, small cell lung cancer

Tleukin (F16-IL2)

Extra-domain A1 of tenascin-C

Semi-synthetic, scfv

ETH-2

Phase Ⅱb

Merkel cell carcinoma, solid tumour, breast cancer

Tesidolumab (LFG316)

Complement C5

Synthetic, Fab

MorphoSys

Phase Ⅱ

Paroxysmal nocturnal hemoglobinuria, geographic atrophy

Tralokinumab (CAT354/BAK1.1)

Interleukin 13

Naive, scFv BAK1 scFv affinity maturation by phage display

CAT

Phase Ⅲ (currently terminated)

Inadequately controlled asthma

Utomilumab (PF05082566)

4-1BB (CD137, TNFRSF9)

Synthetic, Fab

MorphoSys

Phase Ⅲ

Diffuse large B-Cell lymphoma

Vantictumab (OMP18R5)

Frizzled family receptor 7 (Fzd7)

Synthetic, Fab

MorphoSys

Phase Ⅰ

Metastatic breast cancer, solid tumors

VAY-736

B-cell-activating factor receptor (BAFF-R)

Synthetic, Fab

MorphoSys

Phase Ⅲ

Autoimmune hepatitis

Guselkumab Tremfya®

P19 subuntit of interleukin 23 synthetic

MorphoSys, Fab

Morphosys

FDA approved 2017

Pustular psoriasis