Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain

V. Ghetie; M. A. Till; M. A. Ghetie; J. W. Uhr; E. S. Vitetta

doi:10.1016/0022-1759(90)90021-M

Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain

V. Ghetie, M. A. Till, M. A. Ghetie, J. W. Uhr, E. S. Vitetta

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-α-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected HIV-1.

Original language	English (US)
Pages (from-to)	135-141
Number of pages	7
Journal	Journal of Immunological Methods
Volume	126
Issue number	1
DOIs	https://doi.org/10.1016/0022-1759(90)90021-M
State	Published - Jan 24 1990

Keywords

CD4
Human immunodeficiency virus
Immunoconjugate
Ricin A chain

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/0022-1759(90)90021-M

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title = "Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain",

abstract = "A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-α-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected HIV-1.",

keywords = "CD4, Human immunodeficiency virus, Immunoconjugate, Ricin A chain",

author = "V. Ghetie and Till, {M. A.} and Ghetie, {M. A.} and Uhr, {J. W.} and Vitetta, {E. S.}",

note = "Funding Information: ology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75235, U.S.A. • This work is supported by NIH Grants nos. CA-28149, CA-41081 and agrant from theWelch Foundation(no. 1-0947).",

year = "1990",

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doi = "10.1016/0022-1759(90)90021-M",

language = "English (US)",

volume = "126",

pages = "135--141",

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T1 - Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain

AU - Ghetie, V.

AU - Till, M. A.

AU - Ghetie, M. A.

AU - Uhr, J. W.

AU - Vitetta, E. S.

N1 - Funding Information: ology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75235, U.S.A. • This work is supported by NIH Grants nos. CA-28149, CA-41081 and agrant from theWelch Foundation(no. 1-0947).

PY - 1990/1/24

Y1 - 1990/1/24

N2 - A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-α-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected HIV-1.

AB - A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-α-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected HIV-1.

KW - CD4

KW - Human immunodeficiency virus

KW - Immunoconjugate

KW - Ricin A chain

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ER -

Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain

Abstract

Keywords

ASJC Scopus subject areas

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