[Preparation of monoclonal antibodies against HA protein of subtype H1 of swine influenza virus]

Abstract

Aim: To prepare the monoclonal antibodies(mAb) against the HA protein of subtype H1 of swine influenza virus (SIV).

Methods: To construct recombinant expression plasmid subtype H1 of SIV HA gene of A/Swine/Guangdong/LM/2004(H1N1).BALB/c mice of 6-8 weeks old were immunized endemically with the recombinant plasmid.The splenocytes from the immunized mice were fused with Sp2/0 myeloma cells after the last immunization. Hybridoma cells were screened by ELISA and hemagglutination inhibition (HI) tests.The activity and specificity of mAbs were evaluated by HI test and Western blot assay.

Results: Three hybridoma cell lines secreting specific mAbs named 8C4, 8C6, and 9D6 were developed. The ELISA titer of these mAbs was 1:16 000-1:256 000, the HI titer was 1:512-1:256 000; The neutralized titer of 8C4, 8C6, and 9D6 to A/Swine/Guangdong/LM/2004(H1N1) was 10(-2.83), 10(-6.4) and 10(-5.8). Western blot analysis confirmed that mAbs only reacted with HA protein of subtype H1 of SIV.

Conclusion: These mAbs can be used as a useful tool to analyze the antigenic variation.They also provide the effective reagents for the rapid detection of subtype H1 of SIV.