Preparation and cDNA sequence analysis of two novel monoclonal antibodies against magaininII

By using intrasplenic immunization and the conventional B lymphocyte hybridoma technique, we have established two novel hybridoma cell lines stably secreting specific monoclonal antibodies (MAbs) to magaininII, termed as 2D1 and 3F8, respectively, The two cell lines were then subjected to RNA extraction and the V-H and V-L segments were obtained by reverse transcription of RNA followed by polymerase chain reaction (RT-PCR) and characterized by nucleotide sequence analysis. The V-H segments of 2D1 and 3F8 belong to the VH5 family and the V-L segments of 2D1 and 3F8 belong to V(K)10 and V(K)1 groups, respectively. The two MAbs utilize different V-L segments and have disparities in their HCDR3 regions, which may contribute to the different epitope recognition of the two antibodies.