[Preparation of monoclonal antibodies against human Apo B₁₀₀ and establishment of a sandwich ELISA system].

AIM: To prepare the monoclonal antibodies (mAbs) against human Apo B₁₀₀ and establish a double-antibody sandwich ELISA system for human Apo B₁₀₀ detection.

METHODS

The BALB/c mice were immunized with human Apo B₁₀₀ antigen. After cell fusion and screening, the hybridoma cells were cultured in serum-free medium and the supernatant was purified to obtain mAbs using protein A. The affinity, subtype, specificity and epitope of the mAbs were characterized and the sandwich ELISA system was established.

RESULTS

Four hybridoma cell lines 4-1-2, 4-2-2, 4-3-2 and 4-6-3 were obtained. The affinity of the anti-Apo B₁₀₀ mAbs was up to 1×10⁹ L/mol and nearly had no cross reaction with other relevant proteins. Linear detection of the sandwich ELISA system using 4-3-2 and 4-6-3 covered a range from (1.3-80) ng/mL, and its sensitivity was 1.24 ng/mL. The intra-assay coefficient of variation (CV) and inter-assay CV were less than 10% and 15%, respectively, and the recovery rate was above 90%.

CONCLUSION

The mAbs against human Apo B₁₀₀ have been prepared and a sandwich ELISA system for human Apo B₁₀₀ detection has been established successfully, which provide a basis for human Apo B₁₀₀ detection and disease diagnosis.