Full
Outline
To obtain monoclonal antibodies of the factor H binding protein (fHBP) A and B subfamily proteins in meningococcal group B, and establish an ELISA detection method which can provide a new way for quantitation and identification of fHBP antigen.
Mice were immunized with purified histidine-labeled recombinant fHBP (His), the spleen cells of mouse were fused with SP2/0 cells. 3 cell strains with stable secretion of anti-fHBP monoclonal antibodies were acquired, respectively named A, B and AB. To establish a double-antibody sandwich ELISA method for detecting recombinant fHBP antigen, AB was coated as primary antibodies, A and B were labeled by horseradish peroxidase (HRP) as the second antibody.
The GMTs valences of A, B and AB antibodies produced in mouse ascites were all between 4.4~8.8×105, and Western Blot showed that the 3 antibodies could specifically bind the corresponding recombinant fHBP antigens. The direct ELISA test also demonstrated that the 3 monoclonal antibodies merely reacted with proteins containing fHBP A or fHBP B through different recognition sites on antigens, independent of each other. The double-antibody sandwich ELISA method had good specificity and applicability, with sensitivity up to 10 ng·mL-1 and coefficient variation of less than 10%.
The double antibody sandwich ELISA method based on monoclonal antibody has good specificity and applicability, which laid a foundation to further establish a sensitive and rapid method for detection and identification of fHBP antigen.
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| 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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