Chemistry: analytical and immunological testing – Involving producing or treating antigen or hapten
Patent
1984-01-19
1988-02-16
Warden, Robert J.
Chemistry: analytical and immunological testing
Involving producing or treating antigen or hapten
436519, 424 88, 536123, 536124, 514 54, 530387, 530411, 530813, G01N 33531
Patent
active
047255579
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to techniques for producing fucosyl antigens and antibodies for recognizing the same, and more particularly to a process for producing carbohydrate antigens containing a specific carbohydrate linkage as a hapten and a process for producing from the antigens antibodies having specific reactivity with cells of cancers of the digestive system, especially human colon carcinoma cells, and murine teratocarcinoma cells. The present invention also relates to a technique for determining cancer associated carbohydrate linkages with use of such antibodies capable of specifically recognizing specific carbohydrate linkages.
At certain stages of cellular differentiation, specific carbohydrate antigens are recently expressed on the surface of mammalian cells. Monoclonal antibodies [Cell, Vol. 14, 775-783 (1978), Proc., Natl., Acad., USA, Vol. 75, No.11, 5565-5569 (1978) and Nature, Vol. 292, 156-158 (1981)] produced by the hybridoma technique using whole cells as the immunogen, and the antibodies present in sera of some patients [Exp. Cell Res., 131, 185-195 (1981)] have been proposed as antibodies which are reactive with such carbohydrate antigens. In the course of our research conducted in connection with the abovementioned reports, we have succeeded in preparing carbohydrate antigens from specific carbohydrate linkages obtained by organic synthesis and used as haptens and also in producing from the carbohydrate antigens antibodies which specifically selectively react with cells of cancers of the digestive system, especially human colon carcinoma cells, and murine teratocarcinoma cells. We have further obtained the novel finding that the antibodies are useful for recognizing and determining carcinoma cells and for diagnosing cancers. The present invention has been accomplished based on these novel findings.
DESCRIPTION OF THE INVENTION
The present invention provides a process for producing a fucosyl antigen characterized in that an oligosaccharide containing an .alpha.-fucopyranosyl-(1.fwdarw.3)-, -(1.fwdarw.4)- or -(1.fwdarw.6)-galactopyranosyl group and serving as a hapten is reacted with a carrier protein to obtain a carbohydrate antigen.
The invention further provides a process for producing an antibody for recognizing the fucosyl antigen characterized by administering to a mammal a carbohydrate antigen comprising a complex of a carrier protein and an oligosaccharide containing an .alpha.-fucopyranosyl-(1.fwdarw.3)-, -(1.fwdarw.4)- or -(1.fwdarw.6)-galactopyranosyl group, and collecting the resulting antibody.
The invention further provides a method of determining a cancer associated carbohydrate linkage having an .alpha.-fucopyranosyl-(1.fwdarw.3)-, -(1.fwdarw.4)- or -(1.fwdarw.6)galactopyranosyl group characterized in that the carbohydrate linkage is determined by a immunoreaction with use of an antibody capable of recognizing the group specifically.
The invention further provides a cancer diagnosing kit containing an antibody capable of specifically recognizing the .alpha.-fucopyranosyl-(1.fwdarw.3)-, -(1.fwdarw.4)- or -(1.fwdarw.6)-galactopyranosyl group.
Hereinafter described are the production of fucosyl antigens, production of antibodies from the antigens, cancer diagnosing kit containing the antibody, and determination of cancer associated carbohydrate linkages or cancer diagnosis method with use of the antibody, all according to the present invention.
In producing fucosyl antigens according to the present invention, it is essential to use as haptens oligosaccharides containing an .alpha.-fucopyranosyl(1.fwdarw.3)-, -(1.fwdarw.4)- or -(1.fwdarw.6)-galactopyranosyl group. The fucopyranose and galactopyranose, which are essential component sugars of the oligosaccharide, are linked together by .alpha.1-3, .alpha.1-4 or .alpha.1-6 bond, of which the .alpha.1-3 bond is especially preferred. The galactopyranosyl group of the oligosaccharide may have attached thereto another carbohydrate linkage. Typical of sugars constituting such linkages is, for example, glucopy
REFERENCES:
patent: 4078049 (1970-03-01), Felix et al.
patent: 4238473 (1980-12-01), Lemieux et al.
Chiba Taku
Miyauchi Teruo
Muramatsu Takashi
Ozawa Masayuki
Sato Eiichi
Otsuka Pharmaceutical Co. Ltd.
Warden Robert J.
Wieder Stephen C.
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