Large scale preparation of immunotoxins constructed with the Fab′ fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain

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

doi:10.1016/0022-1759(88)90367-5

Large scale preparation of immunotoxins constructed with the Fab′ fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain

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

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Abstract

In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab′ fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab′ and dgA chain and the purification of the Fab′-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab′-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.

Original language	English (US)
Pages (from-to)	267-277
Number of pages	11
Journal	Journal of Immunological Methods
Volume	112
Issue number	2
DOIs	https://doi.org/10.1016/0022-1759(88)90367-5
State	Published - Sep 13 1988

Keywords

Fab′ fragment
Immunotoxin
Large scale preparation of proteins
Monoclonal antibody
Ricin toxin deglycosylated A chain

ASJC Scopus subject areas

Immunology and Allergy
Immunology

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10.1016/0022-1759(88)90367-5

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title = "Large scale preparation of immunotoxins constructed with the Fab′ fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain",

abstract = "In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab′ fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab′ and dgA chain and the purification of the Fab′-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab′-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.",

keywords = "Fab′ fragment, Immunotoxin, Large scale preparation of proteins, Monoclonal antibody, Ricin toxin deglycosylated A chain",

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

note = "Funding Information: Correspondence to: E.S. Vitetta, Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235, U.S.A. * Supported by NIH Grants no. CA-28149 and no. CA-41081, and a grant from the Welch Foundation (no. 1-0947).",

year = "1988",

month = sep,

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doi = "10.1016/0022-1759(88)90367-5",

language = "English (US)",

volume = "112",

pages = "267--277",

journal = "Journal of Immunological Methods",

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T1 - Large scale preparation of immunotoxins constructed with the Fab′ fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain

AU - Ghetie, V.

AU - Ghetie, M. A.

AU - Uhr, J. W.

AU - Vitetta, E. S.

N1 - Funding Information: Correspondence to: E.S. Vitetta, Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75235, U.S.A. * Supported by NIH Grants no. CA-28149 and no. CA-41081, and a grant from the Welch Foundation (no. 1-0947).

PY - 1988/9/13

Y1 - 1988/9/13

N2 - In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab′ fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab′ and dgA chain and the purification of the Fab′-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab′-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.

AB - In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab′ fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab′ and dgA chain and the purification of the Fab′-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab′-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.

KW - Fab′ fragment

KW - Immunotoxin

KW - Large scale preparation of proteins

KW - Monoclonal antibody

KW - Ricin toxin deglycosylated A chain

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JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 2

ER -

Large scale preparation of immunotoxins constructed with the Fab′ fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain

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Keywords

ASJC Scopus subject areas

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