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A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor.

Research output: Contribution to Journal/MagazineJournal article

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A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor. / Kurucz, I.; Jost, C. R.; George, A. J. T. et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 9, 1993, p. 3830-3834.

Research output: Contribution to Journal/MagazineJournal article

Harvard

Kurucz, I, Jost, CR, George, AJT, Andrew, SM & Segal, DM 1993, 'A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor.', Proceedings of the National Academy of Sciences of the United States of America, vol. 90, no. 9, pp. 3830-3834. <http://www.pnas.org/content/90/9/3830.abstract?sid=cca2352d-9364-4674-870f-986c5855ac81>

APA

Kurucz, I., Jost, C. R., George, A. J. T., Andrew, S. M., & Segal, D. M. (1993). A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor. Proceedings of the National Academy of Sciences of the United States of America, 90(9), 3830-3834. http://www.pnas.org/content/90/9/3830.abstract?sid=cca2352d-9364-4674-870f-986c5855ac81

Vancouver

Kurucz I, Jost CR, George AJT, Andrew SM, Segal DM. A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor. Proceedings of the National Academy of Sciences of the United States of America. 1993;90(9):3830-3834.

Author

Kurucz, I. ; Jost, C. R. ; George, A. J. T. et al. / A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor. In: Proceedings of the National Academy of Sciences of the United States of America. 1993 ; Vol. 90, No. 9. pp. 3830-3834.

Bibtex

@article{cc10792c4e3c4e25ba96b9b81e51039a,
title = "A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor.",
abstract = "A single-chain Fv construct of the 2B4 T-cell receptor has been made and expressed in Escherichia coli as bacterial inclusion bodies. After solubilization in 6 M guanidine hydrochloride and formation of mixed disulfides with glutathione, the protein was refolded by diluting out the denaturant and allowing intramolecular disulfide bridges to form by disulfide exchange. Approximately 65-100 mg of refolded protein was obtained from 1 liter of bacterial culture, an appreciable fraction of which was monomeric in nondenaturing solvents. This protein bound to three monoclonal antibodies specific for allotypic or idiotypic determinants on the native 2B4 variable region but did not bind several other anti-T-cell-receptor monoclonal antibodies that lacked such specificity. These experiments show that T-cell-receptor variable regions, like the V regions of antibodies, can form a well-behaved single-chain Fv molecule and provide large amounts of recombinant single-chain Fv T-cell receptor that can be used to study T-cell function.",
author = "I. Kurucz and Jost, {C. R.} and George, {A. J. T.} and Andrew, {S. M.} and Segal, {D. M.}",
year = "1993",
language = "English",
volume = "90",
pages = "3830--3834",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "9",

}

RIS

TY - JOUR

T1 - A bacterially expressed single-chain Fv construct from the 2B4 T-cell receptor.

AU - Kurucz, I.

AU - Jost, C. R.

AU - George, A. J. T.

AU - Andrew, S. M.

AU - Segal, D. M.

PY - 1993

Y1 - 1993

N2 - A single-chain Fv construct of the 2B4 T-cell receptor has been made and expressed in Escherichia coli as bacterial inclusion bodies. After solubilization in 6 M guanidine hydrochloride and formation of mixed disulfides with glutathione, the protein was refolded by diluting out the denaturant and allowing intramolecular disulfide bridges to form by disulfide exchange. Approximately 65-100 mg of refolded protein was obtained from 1 liter of bacterial culture, an appreciable fraction of which was monomeric in nondenaturing solvents. This protein bound to three monoclonal antibodies specific for allotypic or idiotypic determinants on the native 2B4 variable region but did not bind several other anti-T-cell-receptor monoclonal antibodies that lacked such specificity. These experiments show that T-cell-receptor variable regions, like the V regions of antibodies, can form a well-behaved single-chain Fv molecule and provide large amounts of recombinant single-chain Fv T-cell receptor that can be used to study T-cell function.

AB - A single-chain Fv construct of the 2B4 T-cell receptor has been made and expressed in Escherichia coli as bacterial inclusion bodies. After solubilization in 6 M guanidine hydrochloride and formation of mixed disulfides with glutathione, the protein was refolded by diluting out the denaturant and allowing intramolecular disulfide bridges to form by disulfide exchange. Approximately 65-100 mg of refolded protein was obtained from 1 liter of bacterial culture, an appreciable fraction of which was monomeric in nondenaturing solvents. This protein bound to three monoclonal antibodies specific for allotypic or idiotypic determinants on the native 2B4 variable region but did not bind several other anti-T-cell-receptor monoclonal antibodies that lacked such specificity. These experiments show that T-cell-receptor variable regions, like the V regions of antibodies, can form a well-behaved single-chain Fv molecule and provide large amounts of recombinant single-chain Fv T-cell receptor that can be used to study T-cell function.

M3 - Journal article

VL - 90

SP - 3830

EP - 3834

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 9

ER -