Biomedical subjects

C D Surh

Publications and source records attributed to C D Surh.

57 records · Page 4Linked to original sources

PubMed1985

Immunodetection of cell-bound antigens using both mouse and human monoclonal antibodies.

A micro enzyme-linked immunoassay (EIA) has been developed for the rapid and sensitive detection of either human or mouse monoclonal antibodies reactive with cell bound antigens. Whole intact cells are immobilized onto 96-well flat bottom microtiter plates by drying in an oven at 37 degrees C overnight prior to the start of the assay. This method of attachment was suitable for all cell types tested, regardless of origin, size and chromosomal content. The dried cells were then rehydrated, incubated with the appropriate test hybridoma supernatant, followed with subsequent analysis by EIA. The plates can be stored at 4 degrees C up to 1 month for future EIA analysis. This assay offers high sensitivity, requires only small amounts of target cells and test hybridoma supernatant, and can be completed within 3 h. This EIA is well suited for the rapid screening of large numbers of hybridoma supernatants and can also be adapted to include cells of any species, providing the appropriate antibody reagents are available.

Animals↗

PubMed1984

Murine monoclonal antibodies that identify antigenically distinct subpopulations of human sperm.

Murine monoclonal antibodies (MoAbs) were isolated to characterize antigenically distinct subpopulations of human sperm. Spleen cells from Balb/c mice immunized with freshly prepared human sperm, were fused with murine P3-NS1-Ag4-1 myeloma cells by somatic cell hybridization, and supernatants from the IgG-secreting hybridomas were screened by an enzyme immunoassay (EIA) for reactivity against fresh human sperm and a panel of human somatic cells. Two MoAbs, SP1D1 and SP7A7, reacted specifically with human sperm, whereas three others, SP2A9, SP3B3, and SP4F5, cross-reacted with a variety of human somatic cells. The binding of MoAbs were characterized by immunofluorescence, agglutination, and Staphylococcus aureus binding assays. We found that certain MoAbs bound to common antigens of the head and tail, or to tail alone, and had agglutinating activity. However, not all sperm were reactive to antibody, and the binding activity could only be demonstrated in subpopulations of sperm ranging from 5 to 50% of the total number.

Agglutination Tests↗

PubMedDate not supplied

The predominance of IgG3 and IgM isotype antimitochondrial autoantibodies against recombinant fused mitochondrial polypeptide in patients with primary biliary cirrhosis.

Autoantibodies against inner mitochondrial membrane proteins are a hallmark of primary biliary cirrhosis. Specifically, these antimitochondrial autoantibodies recognize two polypeptides of approximately 70 and 52 kD, respectively. Although the specificity of antimitochondrial autoantibodies has been studied for the past 2 decades, the complementary DNA encoding the major primary biliary cirrhosis-specific 70 kD antigen has only recently been cloned. The availability of the recombinant autoantigen has resulted in the development of a highly sensitive and specific ELISA to detect antimitochondrial autoantibodies and to determine their immunoglobulin isotypes. We report herein that IgG3 is the predominant isotype of antimitochondrial autoantibodies in a group of 74 primary biliary cirrhosis patients. This finding is significant in light of the genomic immunoglobulin in heavy chain gene arrangement. Ninety-three per cent of primary biliary cirrhosis patients possessed IgG3 antimitochondrial autoantibodies with titers of 1:10(3) or higher; 32% of these patients possessed titers of 10(4), 29% at 10(5) and 7% at 10(6). IgM antimitochondrial autoantibodies were next most prevalent; 63% of the patients were positive and 50% of these patients showed titers of 10(3), 43% at 10(4) and 6% at 10(5). Other isotypes were present but in much lower titer and occurrence. Isotypes of antimitochondrial autoantibodies reactive to the 52 kD antigen were also determined using immunoblotting techniques. The predominance of IgG3 and IgM were similarly observed. Finally, the serum immunoglobulin isotype levels of primary biliary cirrhosis patients were compared with healthy normal adults by radial immunodiffusion. Serum IgG3 and IgM were very elevated in primary biliary cirrhosis; with IgG3 at 5.5-fold and IgM at 4.3-fold above normals.(ABSTRACT TRUNCATED AT 250 WORDS)

Autoantibodies↗