A unique type of combinatorial protein libraries has been constructed. These libraries are based on the pre-B cell receptor (pre-BCR). The pre-BCR is a protein that is produced during normal development of the antibody repertoire. Unlike that of canonical antibodies, the pre-BCR subunit is a trimer that is composed of an antibody heavy chain paired with two surrogate light chain (SLC) component...

Efficient generation of catalytic antibodies is uniquely dependent on the exact nature of the binding interactions in the antigen-antibody complex. Current methods for generation of monoclonal antibodies do not efficiently survey the immunological repertoire and, therefore, they limit the number of catalysts that can be obtained. We are exploring methods to clone and express the immunological r...

Psychotic episodes in lupus patients can be triggered by an antibody attack on brain cells, according to Matus et al. (page 3221). The psychosis that occurs in a small subset of lupus patients is associated with an antibody that recognizes a trio of ribosomal phosphoproteins. It was assumed that this antibody binds to the cell surface version of one of the phosphoproteins and somehow penetrates...

The variable-region genes of monoclonal antibody against Bacillus cereus spores were cloned from mouse hybridoma cells by reverse transcription-PCR. The heavy- and light-chain variable-region genes were connected by a 45-base linker DNA to allow folding of the fusion protein into a functional tertiary structure. For detection of protein expression, a 10-amino-acid strep tag (biotin-like peptide...

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...