Immunologic compositions methods of preparation and use

Claim 1

An immunologic composition comprising pyruvic aldehyde stabilized erythrocytes sensitized with a polypeptide or glycoprotein antigen selected from the group consisting of chorionic gonadotrophin, pregnant mare's serum gonadotrophin, carcino embryonic antigen, luteinizing hormone, follicle stimulating hormone, human menopausal gonadotrophin and thyroid stimulating hormone, said stabilized erythrocytes being coupled to said antigen with a bifunctional molecule selected from the group consisting of glutaraldehyde, glyoxal, succinaldehyde, hexamethylene diisocyanate, toluene 2,4-diisocyanate, dimethyl suberimidate.

Claim 12

A method for preparing immunologic compositions of animal erythrocytes sensitized with antigens useful in passive haemagglutination tests which comprises: (a) collecting animal blood directly into Alsevers solution so that a final ratio of blood to Alsevers solution is 1:1 in this mixture and wherein the erythrocyte concentration packed cell volume of said blood is from about 20% to about 25%; (b) separating the erythrocytes from said mixture by centrifugation; (c) washing said erythrocytes with normal saline and then suspending said erythrocytes in normal saline at a concentration of about 20%; (d) mixing the suspension of erythrocytes with a buffered mixture of pyruvic aldehyde and normal saline, wherein said pyruvic aldehyde-erythrocyte mixture is phosphate buffered to pH 8.0; (e) incubating the stabilization mixture of step (d) at 37° C. for 3 hours, and collecting the stabilized erythrocytes by centrifugation followed by washing with normal saline; (f) suspending the stabilized erythrocytes in a phosphate buffered solution of antigen; (g) forming a sensitization mixture by thoroughly agitation the suspension of stabilized erythrocytes-antigen with a solution of a bifunctional molecule selected from the group consisting of glutaraldehyde, glyoxal, succinaldehyde, hexamethylene diisocyanate, toluene 2,4-diisocyanate and dimethyl suberimidate in saline; (h) mixing the sensitization mixture for a further 2 hours at room temperature; (i) collecting the sensitized erythrocytes by centrifugation and subsequently washing them with phosphate buffered saline; (j) suspending the sensitized erythrocytes in phosphate buffered saline containing normal rabbit serum wherein the complement of said rabbit serum had previously been fixed.

Claim 13

A method for preparing immunologic compositions of animal erythrocytes sensitized with antigens useful in passive haemagglutination tests which comprises: (a) collecting animal blood directly into Alsevers solution so that a final ratio of blood to Alsevers solution is 1:1 in this mixture and wherein the erythrocyte concentration packed cell volume of said blood is from about 20% to about 25%; (b) separating the erythrocytes from said mixture by centrifugation; (c) washing said erythrocytes with normal saline and then suspending said erythrocytes in normal saline at a concentration of about 20%; (d) mixing the suspension of erythrocytes with a buffered mixture of pyruvic aldehyde and normal saline, wherein said pyruvic aldehyde-erythrocyte mixture is phosphate buffered to pH 8.0; (e) incubating the stabilization mixture of step (d) at 37° C. for 3 hours, and collecting the stabilized erythrocytes by centrifugation followed by washing with normal saline; (f) suspending the stabilized erythrocytes in a phosphate buffered solution of antigen; (g) forming a sensitization mixture by thoroughly agitation the suspension of stabilized erythrocytes-antigen with a solution of a bifunctional molecule selected from the group consisting of glutaraldehyde, glyoxal, succinaldehyde, hexamethylene diisocyanate, toluene 2,4-diisocyanate and dimethyl suberimidate in saline; (h) mixing the sensitization mixture for a further 2 hours at room temperature; (i) collecting the sensitized erythrocytes by centrifugation and subsequently washing them with phosphate buffered saline; (j) suspending the sensitized erythrocytes in phosphate buffered saline containing normal rabbit serum wherein the complement of said rabbit serum had previously been fixed.