Premium
Isolation of a soluble tumor‐associated antigen from human melanoma
Author(s) -
Roth Jack A.,
Holmes E. Carmack,
Reisfeld Ralph A.,
Slocum Harry K.,
Morton Donald L.
Publication year - 1976
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/1097-0142(197601)37:1<104::aid-cncr2820370115>3.0.co;2-i
Subject(s) - antigen , complement fixation test , melanoma , antigenicity , medicine , antibody , delayed hypersensitivity , immunology , cancer research , serology
Human melanoma‐associated antigen was solubilized from fresh surgical specimens by 3 M KCl extraction. The antigenicity of this extract was demonstrated by delayed cutaneous hypersensitivity responses and inhibition of complement fixation. Twenty‐one of 33 melanoma patients had delayed cutaneous hypersensitivity responses to melanoma antigen, while only four of 28 reacted to autologous muscle. Although KCl extracts did not fix complement directly, they reacted with antibody, thus inhibiting complement fixation by the autologous melanoma antigen extracted from tissue culture supernatants. The soluble anti‐genic moiety was then purified by fractionation on a G‐150 Sephedex column and polyacrylamide gels, and the antigenic activity was monitored by delayed cutaneous hypersensitivity responses in melanoma patients. A 20‐fold purification was achieved. Solubilization of tumor antigens with 3 M KCl provides tumor‐associated antigen of high activity which is amenable to further biochemical purification.