Preparation and characterization of monoclonal antibody against recombinant human augmenter of liver regeneration

This study aimed to prepare monoclonal antibody(McAb) against recombinant human augmenter of liver regeneration(rhALR),which may be useful in the development of a specific and convenient method to detect hALR.Balb/c mice were immunized with rhALR,and then splenocytes from the immunized mice were collected and fused with the mouse myeloma cell line SP2/0 cells.The hybridoma cells that secreted anti-rhALR protein McAb antibodies were cloned with limited dilution method.The ascites titers and the affinity of the obtained McAbs were determined by indirect ELISA,while the specificity of the McAbs was tested by indirect ELISA and Western blot analysis.Then the immunoglobulin(Ig) subtype was test by IsoQuick strips,the chromosome of the hybridomas was analyzed.As results,one of hybridomas was screened out,designated 2A6.Chromosome analysis revealed that the obtained hybridomas were possessed universal characteristics of the monoclonal hybridoma cells which secreted McAb,the Ig subtype of 2A6 McAb was IgG2b,and the light chain was kappa.We also detected that he ascites titers of 2A6 McAb was 1:106,and the affinity constant of 2A6 McAb was 3.19×107.Indirect ELISA and Western blot identification suggested that the McAbs react specifically with rhALR protein.In our study,one of high titer,specific McAb against rhALR protein has been successfully prepared and primarily identified.