US 5,270,171 A · Cancer-associated SCM-recognition factor, preparation and method of use

Abstract

A cancer recognition factor (SCM factor) useful in the performance of the structuredness of the cytoplasmic matrix (SCM) test has been isolated, purified to substantial homogeneity, and characterized, and methods for its use have been described. The factor is a peptide of at least 9 amino acid residues including a core sequence of 9 amino acid residues having an amphipathicity profile substantially equivalent to that of the sequence F-L-M-I-D-Q-N-T-K and produces at least a 10 percent decrease in the intracellular fluorescence polarization value of SCM-responding lymphocytes from donors afflicted with cancer. A synthetic SCM factor representing a consensus sequence of M-I-P-P-E-V-K-F-N-K-P-F-V-F-L-M-I-D-Q-N-T-K-V-P-L-F-M-G-K is fully active. Antibodies specific for SCM factor are useful in immunoassays that can detect the factor, including detection in cancer cells grown in vitro. The SCM factor is useful for screening of blood samples and other body fluids or cell aspirates for the presence of malignancy in the donor. The multiple action spectrum of the SCM factor including cancer proliferation and invasion promotion, as well as inhibition of the host's immune defense mechanisms and synthesis of SCM factor by cancer cells, represents a novel target for cancer management. Methods for reducing in vivo activity of the SCM factor, such as dialysis or antibody neutralization, can also be useful in the management of cancer.

Claims68

Dependent claims

A substantially purified cancer recognition factor, the factor being a peptide of nine amino acid residues to 35 amino acid residues including a core sequence of nine amino acid residues having an amphipathicity profile substantially equivalent to that of the sequence F-L-M-I-D-O-N-T-K, wherein the sixth amino acid of the core sequence is selected from the group consisting of Q and N, the seventh amino acid of the core sequence is selected from the group consisting of N and Q, and the ninth amino acid of the core sequence is selected from the group consisting of K and R, the factor producing at least a ten percent (10%) decrease in the intracellular fluorescence polarization value of lymphocytes capable of responding in the structuredness of the cytoplasmic matrix (SCM) test as isolated from donors afflicted with cancer.

A substantially purified cancer recognition factor useful in the structuredness of the cytoplasmic matrix (SCM) test consisting essentially of a peptide have an amino acid sequence of M-X.sub.2 -P-P-X.sub.5 -X.sub.6 -K-F-X.sub.9 -K-P-F-X.sub.13 -F-X.sub.15 -M-X.sub.17 -X.sub.18 -X.sub.19 -X.sub.20 -X.sub.21 -K-X.sub.23 -P-X.sub.25 -F-M-G-K, wherein X.sub.2, X.sub.6, X.sub.13, X.sub.15, X.sub.17, X.sub.23 and X.sub.25 are each independently selected from the group consisting of I, L, and V;

X.sub.5 and X.sub.18 are each independently selected from the group consisting of D and E;
X.sub.9, X.sub.19, and X.sub.20 are each independently selected from the group consisting of Q and N;
and X.sub.21 is selected from the group consisting of S and T, the factor producing at least a 10 percent decrease in the intracellular fluorescence polarization value of SCM-responding lymphocytes from donors afflicted with cancer.

A substantially purified cancer recognition factor active in the structuredness of the cytoplasmic matrix (SCM) test consisting essentially of low molecular weight peptide passing through filters with a nominal 1000-dalton molecular weight cutoff and retained by filters with a nominal 500-dalton molecular weight cutoff, the factor being substantially free of intact .alpha..sub.1 -antitrypsin or other molecules larger than about 10,000 daltons, the factor producing at least a 10 percent decrease in the intracellular fluorescence polarization value of SCM-responding lymphocytes from donors afflicted with cancer as measured by the standard SCM test.

A substantially purified cancer factor produced by a process comprising the steps of:

(a) obtaining a body fluid from a donor afflicted with cancer;
(b) separating a first fraction of the body fluid comprising molecules having an apparent molecular weight of greater than 1000 daltons as determined by ultrafiltration through an ultrafilter with a nominal molecular weight cutoff of 1000 daltons, from a second fraction comprising molecules having an apparent molecular weight of less than 1000 daltons;
and
(c) purifying the second fraction to obtain a substantially pure cancer recognition factor, wherein the step of purifying the second fraction comprises the step of desalting the second fraction by loading the second fraction on gel filtration column with a fractionation range of about 0 to about 700 daltons and capable of separating the salts therefrom, eluting the loaded material from the column with water, and collecting that portion eluting at an elution volume between about 0.3 and about 0.5 times the total chromatographic bed volume to increase the specific activity of the cancer recognition factor in the collected desalted portion relative to the specific activity in the second fraction.

1 more element

Dependencies are inferred from the claim text.

Cancer-associated SCM-recognition factor, preparation and method of use

Life of this patent

Cancer-associated SCM-recognition factor, preparation and method of use

Life of this patent

Abstract

Claims68