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

By using intrasplenic immunization and the conventional B lymphocyte hybrid oma technique, we have established two novel hybridoma cell lines stably se creting specific monoclonal antibodies (MAbs) to magaininII, termed as 2D1 and 3F8, respectively, The two cell lines were then subjected to RNA extrac tion 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 nu cleotide sequence analysis. The V-H segments of 2D1 and 3F8 belong to the V H5 family and the V-L segments of 2D1 and 3F8 belong to V(K)10 and V(K)1 gr oups, respectively. The two MAbs utilize different V-L segments and have di sparities in their HCDR3 regions, which may contribute to the different epi tope recognition of the two antibodies.