Development and verification of a de-cross purification method for Neisseria meningitidis group ACYW135 antiserum
10.13200/j.cnki.cjb.004636
- VernacularTitle:A、C、Y、W135群脑膜炎球菌抗血清去交叉纯化方法的建立及验证
- Author:
Ji ZHENG
1
Author Information
1. Zhejiang Yuchang Biotechnology Co., Ltd., Shaoxing 312300, Zhejiang Province, China
- Publication Type:Journal Article
- Keywords:
Neisseria meningitidis(Nm);
Bacterial broth absorption;
Cross-reactivity;
Serum purification;
ELISA
- From:
Chinese Journal of Biologicals
2026;39(01):93-100
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a purification method using bacterial broth absorption to eliminate cross-reactivity in Neisseria meningitidis group ACYW135 antisera, and to verify its applicability for the detection of polysaccharide conjugate vaccines.Methods The New Zealand rabbits were immunized via auricular vein injection with meningococcal serogroups A, C, Y, and W135, respectively, with two rabbits for each group(one male and one female). Immunizations were administered every other day during the first week, totaling three immunizations, followed by continuous immunizations for a total of four weeks to prepare the baseline antisera. Cross-reactivity of the antisera was purified using repeated absorption with heterologous bacterial broth. The cross-reactivity rates between the purified and unpurified antisera and heterologous polysaccharides were compared via ELISA. The specificity of the antisera was further verified by rocket electrophoresis and rate-dependent turbidimetry. The established method was verified for specificity, specificity, reproducibility, robustness and stability.Results Before purification, significant cross-reactivity was observed, with a maximum rate of 16. 25%. After bacterial broth absorption, cross-reactivity rates for all serogroups were reduced to less than 2%, while high geometric mean titers(GMTs) of specific antibodies were maintained. Rocket electrophoresis and rate-dependent turbidimetry confirmed the absence of cross-reactions. The purified serum exhibited good specificity, and had no obvious cross-reaction with other groups of conjugate polysaccharide antigens. The intra-assay CVs of serum detection of each group ranged from 0. 96% to6. 10%, and the inter-assay CVs ranged from 3. 21% to 6. 18%, both less than 15%. The method showed good robustness with the relative error(RE) values less than 8% under varying antigen concentrations(5-15 μg/mL). The titers of the antisera remained stable over a 360-day monitoring period with no significant difference.Conclusion The established bacterial broth absorption method can effectively remove cross-reactivity in Neisseria meningitidis group ACYW135 antisera. The resulting antisera exhibit high specificity, good precision and stability, making them suitable key detection reagents for the quality control of meningococcal polysaccharide conjugate vaccines.
