Monoclonal hybridoma screening by analysis of immunoglobulin light chains

Talbot, Brian G.; MacLean, Sheila J.; Gibson, D. M.

doi:10.1016/0022-1759(85)90108-5

Cited by 7 publications

(6 citation statements)

References 7 publications

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“…After denaturation of the protein extract for 30 min, reduction was carried out for 60 min at room temperature by the addition of 20 pl 1.5% dithiothreitol (DTT). In order to prevent reassociation of the SH groups formed during the reaction, carboxymethylation was performed by adding 20 pl 4% iodoacetamide (BDH, England) or 20 pl4% iodoacetic acid (Sigma) and incubating the mixture in the dark for 60 min at room temperature (Talbot, MacLean & Gibson 1985).…”

Section: Western Blotting a N D Immunodetectionmentioning

confidence: 99%

Characterization of epitopes on the 25 kD protein of the macrogametes/zygotes of Plasmodium falciparum

Fries

Lamers

Smits

et al. 1989

Parasite Immunology

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A sexual stage-specific protein of Plasmodium falciparum with a Mr of 25,000 is one of the target antigens of transmission-blocking antibodies. The contributions of tertiary structure and post-translational modifications (glycosylation and acylation) to the structure of the epitopes on this protein were the subject of detailed investigations. After modification of the three-dimensional structure and modification or cleavage of carbohydrate groups and linked fatty acids, the immunological reactivity was investigated by three different techniques: (i) immunoprecipitation of radiolabelled proteins, (ii) enzyme-linked immunosorbent assay (ELISA), and (iii) Western blotting. The results of the experiments indicate that the immunological reactivity of the major epitopes on the 25 kD protein, including the epitope involved in transmission-blocking immunity, are dependent on the tertiary structure of the protein and on the presence of linked fatty acids, but not on the presence or absence of carbohydrate groups.

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Section: Western Blotting a N D Immunodetectionmentioning

confidence: 99%

Characterization of epitopes on the 25 kD protein of the macrogametes/zygotes of Plasmodium falciparum

Fries

Lamers

Smits

et al. 1989

Parasite Immunology

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“…immunoglobulin light chains [27]. This procedure confirms monoclonality and as sures that each antibody studied will have different properties, although not necessarily different epitopes [33,36].…”

Section: Characterization O F Antijiber Monoclonal Antibodiesmentioning

confidence: 57%

“…The antibodies were analyzed to qualitatively de termine which had different light-chain structures [27], By selecting the antibodies in this way, it is almost certain that each anti body will have different binding characteris tics, if not necessarily different epitopes [36]. Fifteen different antibodies were selected by light-chain analysis.…”

Section: Discussionmentioning

confidence: 99%

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Anti-Idiotype Response to Selected Neutralizing Monoclonal Antibodies Generated against Adenovirus Type 2 Fiber

Talbot

Hallée²

1989

Intervirology

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A series of monoclonal antibodies was prepared against purified adenovirus type 2 fiber. These antibodies were produced by both classical techniques and by electrofusion. Fifty antibodies were obtained, and five were selected which had the following characteristics: (i) they were able to reduce adenovirus 2 infection of Hep-2 cells; (ii) they had different light-chain structures and (iii) they did not react with denatured fiber. The five antibodies (AdF18, AdF38, AdF41, AdF2D3, AdF3Bl) recognized between them three periodate-insensitive epitopes on the fiber. AdF18 bound to both adenovirus type 2 and type 5; all the others were specific to type 2. Polyclonal anti-idiotype antibodies to these antibodies were produced in rabbits and were purified extensively on immunoaffinity columns. Anti-idiotypes to AdF38 and AdF41 were able to inhibit virus infectivity, and anti-AdF41 competed with fiber for binding to the surface of Hep-2 cells.

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“…Antibody cleavage was assessed by acid urea gel electrophoresis (Talbot, 1985). Sample preparation consisted of dialysis against 1 M acetic acid, or addition of acetic acid to 0.1 M. Basic fuchsin was used as tracking dye.…”

Section: Detection Of Immunoglobulin Cleavagementioning

confidence: 99%

A novel acid proteinase relased by hybridoma cells

1990

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An acid proteinase has been detected in culture supernate of the 9.2.27 murine hybridoma. This enzyme extensively degrades albumin and transferrin during short incubations at pH 3 and below. Limited proteolysis of the 9.2.27 IgG2a appears to occur in the culture supernate. Proteolysis in enhanced at low pH in the presence of urea or 1 M acetic acid. The proteinase activity accumulates in continuous perfusion, total cell recycle cultures, beginning during exponential growth of the hybridoma. It is destroyed by boiling and blocked by pepstatin, but not by inhibitors of cysteine or serine proteinases or by EDTA. The low pH optimum may distinguish this enzyme from the known rat and mouse aspartic acid proteinases including cathepsin D and cathepsin E.

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Monoclonal hybridoma screening by analysis of immunoglobulin light chains

Cited by 7 publications

References 7 publications

Characterization of epitopes on the 25 kD protein of the macrogametes/zygotes of Plasmodium falciparum

Characterization of epitopes on the 25 kD protein of the macrogametes/zygotes of Plasmodium falciparum

Anti-Idiotype Response to Selected Neutralizing Monoclonal Antibodies Generated against Adenovirus Type 2 Fiber

A novel acid proteinase relased by hybridoma cells

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