2. Production and characterization of Newcastle disease antibody as a reagent to develop a rapid immunodiagnostic test tool

摘要

Aim: This research was conducted to produce and characterize ND antibody as reagent candidate to develop a rapid immunodiagnostic test tool. Materials and Methods: Four New Zealand White rabbits were used in this study and divided into two groups. First group was injected by Sato ND antigen, and second group was injected by genotype VII ND antigen. This study is divided into three steps: (a) ND antibody production, (b) ND antibody purification, and (c) ND antibody characterization. First group was rabbit injected by Sato NDV (5x108.25 egg lethal doses (ELD)₅₀/ml) and second group was injected by genotype VII NDV (5x106.5 ELD₅₀/ml). Antigen induction was performed by subcutaneous administrated for first (day 1) and second (day 14) injection and intravenous administrated for third (day 30) injection. Blood was collected on day 8 after third injection. Results: Antibody production increased on second antigen injection and reached a peak on day 9 after second antigen injection. Sato and genotype VII ND antibody can be produced without adjuvant within 38 days with the highest titer 210. Based on antibody titer data, both antigens induced antibody production in a similar trend. The characterization antibody by SDS-PAGE indicated that molecular weight of immunoglobulin G (IgG) is 154.93 kDa (whole IgG), heavy chain 54.39 kDa, and light chain 27.74 kDa. ND antibodies have specificity to homologous and heterologous NDVs in varying virulence. Conclusion: Sato and genotype VII ND antibodies have been successfully produced within 38 days without adjuvant. Specificity of ND antibodies to NDVs in varying virulence and cross-reaction between Sato ND antibody and genotype VII ND antibody indicates that the characterized ND antibodies can be used as a reagent to develop rapid immunodiagnostic test tools.