A rapid and simple hapten conjugation method for monoclonal antibodies to be used in immunoenzyme single and double staining procedures

Mullink, H.; Boorsma, D. M.; Klein, Jeanette; Oostendorp, Robert A.J.; Henzen‐Logmans, S. C.; Scheper, R.J.

doi:10.1016/0022-1759(87)90128-1

Cited by 8 publications

(2 citation statements)

References 18 publications

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“…Dinitrophenol was conjugated to mouse A20 IgG mAb as previously described [17] . Briefly, 1-fluoro-2,4-dinitrobenzene (DNFB; Sigma Aldrich) was diluted in chloroform to 50 mg/ml.…”

Section: Methodsmentioning

confidence: 99%

A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies

2011

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We demonstrate here a rapid alternative method for the production of functional bi-specific antibodies using the mild reducing agent 2-mercaptoethanesulfonic acid sodium salt (MESNA). Following reduction of a mixture of two monoclonal antibodies with MESNA to break inter heavy chain bonds, this solution is dialysed under oxidising conditions and antibodies are allowed to reform. During this reaction a mixture of antibodies is formed, including parental antibodies and bi-specific antibody. Bi-specific antibodies are purified over two sequential affinity columns. Following purification, bi-specificity of antibodies is determined in enzyme-linked immunosorbent assays and by flow cytometry. Using this redox method we have been successful in producing hybrid and same-species bi-specific antibodies in a time frame of 6–10 working days, making this production method a time saving alternative to the time-consuming traditional heterohybridoma technology for the production of bi-specific antibodies for use in early pilot studies. The use of both rat and mouse IgG antibodies forming a rat/mouse bi-specific antibody as well as producing a pure mouse bi-specific antibody and a pure rat bi-specific antibody demonstrates the flexibility of this production method.

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“…Dinitrophenol was conjugated to mouse A20 IgG mAb as previously described [17] . Briefly, 1-fluoro-2,4-dinitrobenzene (DNFB; Sigma Aldrich) was diluted in chloroform to 50 mg/ml.…”

Section: Methodsmentioning

confidence: 99%

A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies

2011

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“…Oxazolone‐induced colitis and dermatitis in mice have become recognized models in the study of the immunological mechanisms underlying the diseases . Oxazolone is considered a haptenizing agent which penetrates the mucosal or epithelial barrier and triggers an immunological response, mimicking the described diseases . When applied to the skin of hairless mice (hr/hr) in low doses for a period of 3 weeks, mice develop symptoms characteristic for AD to include barrier dysfunction, secretion of IgE, epithelial cell hyperplasia, fibrosis and influx of inflammatory cells into the dermis and epidermis and secretion of Th2‐characterized interleukins .…”

Section: Animal Models: Oxazolone‐induced Inflammatory Diseasesmentioning

confidence: 99%

The route to pathologies in chronic inflammatory diseases characterized by T helper type 2 immune cells

Jovanovic

Siebeck

Gropp

2014

Clinical and Experimental Immunology

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Transport of glutathione conjugates into secretory vesicles is mediated by the multidrug-resistance protein 1

Luyn¹,

Müller²,

Renes³

et al. 1998

Int. J. Cancer

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A rapid and simple hapten conjugation method for monoclonal antibodies to be used in immunoenzyme single and double staining procedures

Cited by 8 publications

References 18 publications

A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies

A Novel Redox Method for Rapid Production of Functional Bi-Specific Antibodies For Use in Early Pilot Studies

The route to pathologies in chronic inflammatory diseases characterized by T helper type 2 immune cells

Transport of glutathione conjugates into secretory vesicles is mediated by the multidrug-resistance protein 1

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