Preparation of Fab fragments from a mouse monoclonal IgM

Jean M. Bidlack; Peter C. Mabie

doi:10.1016/0022-1759(86)90473-4

Preparation of Fab fragments from a mouse monoclonal IgM

Jean M. Bidlack, Peter C. Mabie

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

21 Scopus citations

Abstract

A procedure is described for preparing and purifying Fab fragments from a mouse monoclonal IgM. The IgM was digested with trypsin in the presence of reducing agent, cysteine. The resulting Fab fragments were alkylated with idoacetamide and purified using a Sephacryl S-200 column. The Fab fragments had a molecular weight of 48 000 as determined by SDS polyacrylamide gel electrophoresis and molecular sieve chromatography. This procedure has been successfully used with five different mouse monoclonal IgM. The Fab fragments bound antigen with the same specificity as the whole IgM.

Original language	English (US)
Pages (from-to)	157-162
Number of pages	6
Journal	Journal of Immunological Methods
Volume	91
Issue number	2
DOIs	https://doi.org/10.1016/0022-1759(86)90473-4
State	Published - Jul 24 1986
Externally published	Yes

Keywords

Fab fragment
IgM, mouse
Monoclonal antibody

ASJC Scopus subject areas

Immunology and Allergy
Immunology

Access to Document

10.1016/0022-1759(86)90473-4

Cite this

@article{3e03c26762f440cfb2e91c5959ef5cae,

title = "Preparation of Fab fragments from a mouse monoclonal IgM",

abstract = "A procedure is described for preparing and purifying Fab fragments from a mouse monoclonal IgM. The IgM was digested with trypsin in the presence of reducing agent, cysteine. The resulting Fab fragments were alkylated with idoacetamide and purified using a Sephacryl S-200 column. The Fab fragments had a molecular weight of 48 000 as determined by SDS polyacrylamide gel electrophoresis and molecular sieve chromatography. This procedure has been successfully used with five different mouse monoclonal IgM. The Fab fragments bound antigen with the same specificity as the whole IgM.",

keywords = "Fab fragment, IgM, mouse, Monoclonal antibody",

author = "Bidlack, {Jean M.} and Mabie, {Peter C.}",

note = "Funding Information: The initial successful cleavage of the IgG was obtained using papain (Porter, 1959). The production of Fab fragments from IgG from a number of different species has been summarized (Mage, 1980). As a result of the increased use of monoclonal antibodies from rats and mice, procedures have been described for optimizing the production of Fab and F(ab')2 fragments from the different subclasses of IgG (Lamoyi and Nisonoff, 1983; Parham, 1983; Rousseaux et al., 1983). Fab frag- * This work was supported by USPHS Grant DA03742 and a grant from the Genesee Valley Heart Association. ** To whom correspondence should be addressed.",

year = "1986",

month = jul,

day = "24",

doi = "10.1016/0022-1759(86)90473-4",

language = "English (US)",

volume = "91",

pages = "157--162",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

publisher = "Elsevier",

number = "2",

}

TY - JOUR

T1 - Preparation of Fab fragments from a mouse monoclonal IgM

AU - Bidlack, Jean M.

AU - Mabie, Peter C.

N1 - Funding Information: The initial successful cleavage of the IgG was obtained using papain (Porter, 1959). The production of Fab fragments from IgG from a number of different species has been summarized (Mage, 1980). As a result of the increased use of monoclonal antibodies from rats and mice, procedures have been described for optimizing the production of Fab and F(ab')2 fragments from the different subclasses of IgG (Lamoyi and Nisonoff, 1983; Parham, 1983; Rousseaux et al., 1983). Fab frag- * This work was supported by USPHS Grant DA03742 and a grant from the Genesee Valley Heart Association. ** To whom correspondence should be addressed.

PY - 1986/7/24

Y1 - 1986/7/24

N2 - A procedure is described for preparing and purifying Fab fragments from a mouse monoclonal IgM. The IgM was digested with trypsin in the presence of reducing agent, cysteine. The resulting Fab fragments were alkylated with idoacetamide and purified using a Sephacryl S-200 column. The Fab fragments had a molecular weight of 48 000 as determined by SDS polyacrylamide gel electrophoresis and molecular sieve chromatography. This procedure has been successfully used with five different mouse monoclonal IgM. The Fab fragments bound antigen with the same specificity as the whole IgM.

AB - A procedure is described for preparing and purifying Fab fragments from a mouse monoclonal IgM. The IgM was digested with trypsin in the presence of reducing agent, cysteine. The resulting Fab fragments were alkylated with idoacetamide and purified using a Sephacryl S-200 column. The Fab fragments had a molecular weight of 48 000 as determined by SDS polyacrylamide gel electrophoresis and molecular sieve chromatography. This procedure has been successfully used with five different mouse monoclonal IgM. The Fab fragments bound antigen with the same specificity as the whole IgM.

KW - Fab fragment

KW - IgM, mouse

KW - Monoclonal antibody

UR - http://www.scopus.com/inward/record.url?scp=0022489907&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022489907&partnerID=8YFLogxK

U2 - 10.1016/0022-1759(86)90473-4

DO - 10.1016/0022-1759(86)90473-4

M3 - Article

C2 - 3016096

AN - SCOPUS:0022489907

SN - 0022-1759

VL - 91

SP - 157

EP - 162

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 2

ER -

Preparation of Fab fragments from a mouse monoclonal IgM

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this