Immunodot assay for determining the isotype and light chain type of murine monoclonal antibodies in unconcentrated hybridoma culture supernates

McDougal, J. Steven; Browning, Sandra W.; Kennedy, Susan; Moore, Deborah

doi:10.1016/s0022-1759(83)80001-5

Cited by 47 publications

(8 citation statements)

References 4 publications

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“…The antibody was used as hybridoma supernatant or as ascites fluid after growing the hybridoma cells in Pristane-primed BALB/c mice. By an immunodot assay (McDougal et al, 1983) the antibody D171 was found to be of the IgG1 isotype. IgG was purified by passing the ascites fluid over a Protein A Sepharose column and eluting the adsorbed IgG with 0.1 M-citrate buffer at pH 4.5 (Ey et al, 1978).…”

Section: Viruses and Cellsmentioning

confidence: 97%

Production of a Monoclonal Antibody against an Epitope on HeLa Cells that Is the Functional Poliovirus Binding Site

Nobis

Zibirre

Meyer

et al. 1985

Journal of General Virology

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SUMMARYCell lines of primate origin carry receptors on their plasma membrane which are responsible for the specific binding of poliovirus.This paper describes the isolation and characterization of a monoclonal antibody reacting with the plasma membrane of HeLa ceils. The antibody (D171) was selected for its protection of HeLa cells against the cytopathic effect of poliovirus type 1. This protection was found to extend to all three viral serotypes, while the replication of five other viruses in HeLa cells was not affected. The 125i_labelled purified antibody did not react with cell lines derived from pig, dog or rodents but bound specifically to all lines of human or primate origin. Immunoglobulin or Fab fragments of DI71 prevented the binding of 35S-labelled poliovirus to HeLa cells. Conversely, nearly all binding sites of 125I-labelled D171 immunoglobulins or Fab fragments could be blocked after preincubation of HeLa cells with poliovirus. These results indicate that D 171 recognizes the poliovirus receptor site on different susceptible cells and that practically all D171 binding sites are involved in the specific attachment of poliovirus to the plasma membrane. To determine whether the epitope recognized by D171 could be separated from the receptor for poliovirus, human-mouse cell hybrids were prepared and analysed. In all 40 clones tested, the susceptibility to poliovirus correlated with the binding of D171.

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Section: Viruses and Cellsmentioning

confidence: 97%

Production of a Monoclonal Antibody against an Epitope on HeLa Cells that Is the Functional Poliovirus Binding Site

Nobis

Zibirre

Meyer

et al. 1985

Journal of General Virology

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“…Monoclonal antibodies BRIC 18 (TgG2a), BRIC 68 (IgG2a), BRIC 107 (IgG1) and BRIC 203 (IgGI) were prepared using spleen cells from Balb/c mice immunized with intact human erythrocytes, essentially as described (Parsons et al, 1982). Immunoglobulin (Ig) isotypes were determined by dot-blotting (McDougal et a/., 1983). Ig was purified and Fab fragments were prepared as previously described (Gardner et al, antibody K-k+Kp(a-b + ) (Marsh, 1988;Anstee & Lisowska, 1990).…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

Monoclonal antibodies against Kell glycoprotein: serology, immunochemistry and quantification of antigen sites

Parsons

Gardner

Anstee

1993

Transfusion Medicine

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Monoclonal antibodies BRIC 18, BRIC 68, BRIC 107 and BRIC 203 recognize high-frequency epitopes absent from erythrocytes expressing the Ko phenotype. BRIC 107 has anti-k (K2)-like specificity. BRIC 203 has a unique specificity denoted anti-Kpbc. All four monoclonal antibodies identify an M(r) 95,600 erythrocyte membrane protein by immunoprecipitation from radio-iodinated erythrocytes. In quantitative binding studies using IgG it is estimated that there are from 2000 (BRIC 18) to 4000 (BRIC 68) copies of the Kell glycoprotein per erythrocyte. Using Fab fragments the estimates are in the range 4000 (BRIC 18) to 18,000 (BRIC 68) copies. In competitive binding assays the four epitopes defined by the BRIC monoclonal antibodies fall into two non-overlapping groups. The first group comprises BRIC 18, BRIC 68, BRIC 203 and an antibody (6-22) with anti-K14 specificity. The second group contains BRIC 107 and two further anti-k-like monoclonal antibodies (BS45 and OSK5). The results suggest that the polymorphisms encoded at the K/k and Kpa/Kpb/Kpc loci may be located in two spatially distinct regions of the Kell glycoprotein(s).

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“…Screening for antibodysecreting cells was done by a dot blot method using SDS-solubilized normal red cell ghosts as antigen. BRIC 163 was an lgG2a antibody as determined by the method of McDougal et al [12],…”

Section: Msmhmentioning

confidence: 99%

A Japanese Family with Two Sisters Apparently Homozygous for M^k

Okubo¹,

Daniels²,

Parsons³

et al. 1988

Vox Sanguinis

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Two Japanese sisters with consanguineous parents have M —N — En(a —) Wr(a — b — ) S — s — U— red cells and are therefore apparently homozygous for M^k; the third reported family with members of this genotype. The serum of the proposita (ORCMK) contained anti-En^aTS, anti-En^a FR and possibly anti-Wr^b, whereas the serum of her M^kM^k sister contained no atypical antibodies. Total absence of sialoglycoproteins α and δ from red cell membranes of an M^k homozygote was demonstrated by lactoperoxidase-catalysed radioiodination of accessible tyrosine residues with subsequent SDS polyacrylamide gel electrophoresis and autoradiography, and by use of a monoclonal antibody directed at the cytoplasmic portion of α-sialoglycoprotein.

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Immunodot assay for determining the isotype and light chain type of murine monoclonal antibodies in unconcentrated hybridoma culture supernates

Cited by 47 publications

References 4 publications

Production of a Monoclonal Antibody against an Epitope on HeLa Cells that Is the Functional Poliovirus Binding Site

Production of a Monoclonal Antibody against an Epitope on HeLa Cells that Is the Functional Poliovirus Binding Site

Monoclonal antibodies against Kell glycoprotein: serology, immunochemistry and quantification of antigen sites

A Japanese Family with Two Sisters Apparently Homozygous for M^k

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