A NEW MARKER FOR HUMAN CANCER CELLS. 1. THE Ca ANTIGEN AND THE Cal ANTIBODY

doi:10.1016/S0140-6736(82)91150-3

The Lancet

Volume 320, Issue 8288, 3 July 1982, Pages 1-6

https://doi.org/10.1016/S0140-6736(82)91150-3 Get rights and content

Abstract

In a search for antibodies that might distinguish between malignant and non-malignant cells a panel of matched pairs of hybrid cells produced by fusion of diploid fibroblasts with malignant cells originating from a cervical carcinoma was used as a screen. Each pair consisted of a hybrid in which malignancy was suppressed and a malignant segregant derived from this hybrid. A monoclonal antibody, designated Cal, was found that discriminated absolutely between the hybrids in which malignancy was suppressed and the malignant segregants. This antibody detected an antigen present in the cell membranes of a wide variety of malignant human cell lines but not of diploid human cell strains. The antigen was found in very low concentrations, if at all, in homogenates of normal adult or fetal tissues. It could be immunoprecipitated by the Cal antibody from extracts of malignant cells but not from extracts of non-malignant cells. After reduction, the immunoprecipitated antigen separated in sodium dodecyl sulphate acrylamide gels as two bands with approximate molecular masses of 390 000 and 350 000. These two components had the properties of glycoproteins with a high carbohydrate content; both bound the Cal antibody. The antigenic determinant resisted boiling at 100°C and extraction by a range of organic solvents. The binding of the Cal antibody to the antigen was substantially reduced by treatment of the antigen with neuraminidase, and the antigenic determinant was largely destroyed by certain endoglycosidases and by extensive proteolysis. Pending its further characterisation, this antigen has been called the Ca antigen.

References (25)

Cj Der et al.
Lack of correlation between the decreased expression of cell surface LETS protein and tumorigenicity in human cell hybrids
Cell
(1978)
Cj Der et al.
A tumor-specific membrane phosphoprotein marker in human cell hybrids
Cell
(1981)
J.O'D. McGee et al.
A new marker for human cancer cells 2. Immunohistochemical detection of the CA antigen in human tissues with the Cal antibody
Lancet
(1982)
H. Harris
Cell fusion and the analysis of malignancy. The Croonian Lecture
Proc Roy Soc B
(1971)
H. Harris
The city of Berlin and some modern approaches to the cancer problem
H. Harris
Cell fusion and the analysis of malignancy
Ds Straus et al.
Growth in vitro of tumour cell × fibroblast hybrids in which malignancy is suppressed
J Coll Sci
(1977)
Fm Watt et al.
The distribution of actin cables and microtubules in hybrids between malignant and non-malignant cells and in tumours derived from them
J Cell Sci
(1978)
Ej Stanbridge et al.
Analysis of malignancy in human cells: malignant and transformed phenotypes are under separate genetic control
Proc Nat Acad Sci
(1978)
N. Howell et al.
Noncoordinate expression of SV40-induced transformation and tumorigenicity in mouse cell hybrids
Somat Cell Genet
(1979)

Me Bramwell et al.

An abnormal membrane glycoprotein associated with malignancy in a wide range of different tumours

Proc Roy Soc B

(1978)

Me Bramwell et al.

Some further information about the abnormal membrane glycoprotein associated with malignancy

Proc Roy Soc B

(1978)

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A NEW MARKER FOR HUMAN CANCER CELLS. 1. THE Ca ANTIGEN AND THE Cal ANTIBODY

Abstract

Cell

Cell

Lancet

Cell fusion and the analysis of malignancy. The Croonian Lecture

Proc Roy Soc B

The city of Berlin and some modern approaches to the cancer problem

Cell fusion and the analysis of malignancy

Growth in vitro of tumour cell &times; fibroblast hybrids in which malignancy is suppressed

J Coll Sci

The distribution of actin cables and microtubules in hybrids between malignant and non-malignant cells and in tumours derived from them

J Cell Sci

Analysis of malignancy in human cells: malignant and transformed phenotypes are under separate genetic control

Proc Nat Acad Sci

Noncoordinate expression of SV40-induced transformation and tumorigenicity in mouse cell hybrids

Somat Cell Genet

An abnormal membrane glycoprotein associated with malignancy in a wide range of different tumours

Proc Roy Soc B

Some further information about the abnormal membrane glycoprotein associated with malignancy

Proc Roy Soc B

Growth in vitro of tumour cell × fibroblast hybrids in which malignancy is suppressed