The lower matrix protein (pp65) is a major product of many laboratory strains of cytomegalovirus (CMV). It is thus an integral part of many CMV serological assays based on native antigen. Recombinant fragments of pp65 have previously been investigated for their usefulness in more-defined assays. The latter antigens have, however, failed to develop a positive response with serum samples derived fro

A new simple, reproducible and sensitive ELISA that uses trichloroacetic acid (TCA) for the coating of lipooligosaccharide (LOS), smooth lipopolysaccharide (LPS) or lipid A to the solid phase has been developed. The experimental parameters (temperature of coating, time of coating, antigen concentration and TCA concentration) were evaluated by a complete factorial design (24). As a result of the ev

The design of an in vitro immunization system based on a synthetic heterotope immunogen, which was a peptide containing both T- and B-cell epitopes, that elicited a neutralizing, primary human humoral immune response against the human immunodeficiency virus (HIV-1) is reported here. This heterotope construct contained the major neutralizing B-cell epitope, within the V3 region of glycoprotein 120

Recombinant DNA technology has made it possible to produce specific Fab and scFv antibody (Ab) fragments in prokaryotic host cells. Using vectors designed for periplasmic expression of encoded Ab fragments, we have been studying how the sequence and genetic localization of the light chain (L-chain) variable region gene of a mouse Ab (CB-Nm.1) determined the level of Ab production. The variable reg

We have studied the influence of bacterial host on the secretion of single-chain Fv antibody fragment (scFv), the production of this antibody fragment as intracellular fusion protein, and the effect of chaperonin coexpression on intracellular antibody expression. Seven bacterial strains were transformed with a vector carrying the genes encoding the variable regions of an anti-CEA scFv antibody and

Glycoprotein B (gB, gpUL55) is a major antigen for the induction of neutralizing antibodies against human cytomegalovirus, making it an attractive antigen for active and passive immunoprophylaxis. The immunodominant region on gB is the antigenic domain 1 (AD-1), a complex structure which requires a minimal linear amino acid sequence of more than 75 amino acids (aa 552-635) for antibody binding. We

Flow cytometry has been utilized to evaluate the stability of antibody production by unstable subclones of a human × human × mouse heterohybridoma. Heterogeneity of cell-associated immunoglobulin heavy chain expression was demonstrated in different subclones and an increased frequency of cells containing low levels of heavy chain was found to correlate with low antibody productivity. However, the

The human antibody response to the conserved neutralization-related site on the gp 116 of human cytomegalovirus (HCMV) was investigated in healthy blood donors by the use of synthetic peptides. Anti-HCMV positive sera investigated in ELISA gave a reactivity of 48-56% with the peptide T7-13 (amino acids (aa) 67-86). Though epitope mapping revealed several individual fine specificities within this r

The humoral immune response to human cytomegalovirus (CMV) membrane glycoprotein gp58/116 (gB) has been studied by establishing cell lines producing specific human monoclonal antibodies. These cell lines were generated from peripheral blood lymphocytes obtained from a healthy carrier. Hybridomas producing gp58/116-speciftc antibodies were detected by reactivity to procaryotically expressed protein