Next-generation sequencing of antibody transcripts provides a wealth of data, but the ability to identify function-specific antibodies solely on the basis of sequence has remained elusive. We previously characterized the VRC01 class of antibodies, which target the CD4-binding site on gp120, appear in multiple donors, and broadly neutralize HIV-1. Antibodies of this class have developmental comm...

Evidence has been presented that rabbit homocytotropic antibody prepared against ovalbumin bears allotypic markers of both group a (heavy chain) and group b (light chain). This was shown by specific removal of activity in passive cutaneous anaphylaxis assays by anti-allotype immunoadsorbents. The homocytotropic antibody has properties which indicate that it belongs to a new class of rabbit immu...

We cloned the heavy- and light-chain antibody genes of a human X (humanxmouse) trioma secreting a neutralizing, IgG monoclonal antibody to the G2-protein of Puumala virus. The antibody genes were inserted separately into plasmid transfer vector pIEI-4 such that the genes were under control of the baculovirus immediate early gene promoter, IEI. Trichoplusia ni (TN) cells were co-transfected with...

In a previous report we presented evidence which indicated that antibody diversity is generated in large part by somatic processes [I] . That evidence was based on DNA-RNA hybridization studies with a purified mouse a-chain mRNA. The present report deals with similar studies using a preparation of mRNA coding for a mouse heavy chain. The results confirm and extend our previous finding, namely t...

In this Application Note, separation matrices for protein characterization by capillary gel electrophoresis (CGE) with UV detection from two suppliers, Beckman Coulter and Advanced Analytical, were compared on the Agilent 7100 CE system. Two different sample sets were analyzed: fi rst, a protein size standard and BSA as a test protein for molecular weight determination; second, a reduced antibo...

Oxidation of methionine (Met) residues is one of several chemical degradation pathways for recombinant IgG1 antibodies. Studies using several methodologies have indicated that Met oxidation in the constant IgG1 domains affects in vitro interaction with human neonatal Fc (huFcRn) receptor, which is important for antibody half-life. Here, a completely new approach to investigating the effect of o...

background: trastuzumab (herceptin) is a humanized monoclonal antibody (mab) which is used for specific treatment of metastatic breast cancer in patients with overexpression of her2/neu receptor. in this study, we have attempted to develop a biosimilar version of trastuzumab mab. methods: according to in silico studies, the heavy and light chains of trastuzumab mab were designed and constructed...

We characterized nine monoclonal antibodies that bind to the heavy chain of Acanthamoeba myosin-IA. Eight of these antibodies bind to myosin-IB and eight cross-react with Acanthamoeba myosin-II. All but one of the antibodies bind to a 30-kD chymotryptic peptide of myosin-IA that derives from the COOH terminus of the molecule, and to tryptic peptides as small as 17 kD, hence these epitopes are c...

Formation of a complete immunoglobulin heavy-chain transcription unit involves the ordered rearrangement of variable (V), diversity (D), and joining (J) region gene segments. In antibody-producing cells, this process is regulated such that only one of two antibody genes is expressed. Experiments with transgenic mice suggest that this mechanism, known as allelic exclusion, is mediated through th...

The heavy- and light-chain variable regions from a murine monoclonal antibody that recognize Pseudomonas aeruginosa serogroup O6 lipopolysaccharide (LPS) were used to generate a series of chimeric mouse-human monoclonal antibodies with identical variable regions. The murine variable-region gene segments were cloned into an immunoglobulin (Ig) cDNA expression vector that contained the human kapp...