DIRECT RADIOIMMUNOASSAY FOR ANTIGENS AND THEIR ANTIBODIES

• US 3,867,517 A
• Filed: 12/21/1971
• Issued: 02/18/1975
• Est. Priority Date: 12/21/1971
• Status: Expired due to Term

• Current Assignee: Chung-Mei Ling
• Original Assignee: Chung-Mei Ling
• Inventors: Ling, Chung-Mei
• Primary Examiner(s): Sebastian, Leland A.
• Application Number:
  US05/210,510
• Time in Patent Office: 1,155 Days
• Field of Search: 424/1 23/23B 250/16T
• US Class Current: 436/531
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/576 for hepatitis
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/82 Hepatitis associated antigens and antibodies

Abstract

A method for direct radioimmunoassay of antigens or their associated antibodies utilizing a coated test well comprising adding a serum to be tested for an antigen to test apparatus coated with its antibody; incubating the tubes for from 0.5 to 42 hours; aspirating the contents and washing the same with a TrisHCl and sodium azide mixture; adding purified I-125 labeled antibody into the tube and incubating for from 1 to 24 hours; aspirating and washing the contents; and counting the tube for I125 radiation. A similar assay for the antibody may be conducted utilizing an antigen coated tube and I-125 labeled antigen. The method for coating these tubes comprises adding the antigen or antibody solution in a Tris-HCl, sodium azide solution into a suitable test tube or well, incubating the tubes at room temperature from 6 to 72 hours, aspirating and washing the contents and storing at from between 2 and 25*C. until use.

Claims

1. A METHOD FOR DETERMINING THE PRESENCE OF A HEPATITIS ASSOCIATED ANTIGEN OR ITS ANTIBODY CAPABLE OF BEING BOUND TO TWO HEPATITIS ASSOCIATED ANTIGENS OR ANTIBODIES RESPECTIVELY IN AN UNKNOWN SAMPLE UTILIZING DIRECT RADIOMMUNOASSAY COMPRISING:

A. FORMING A SOLUTION OF SAID HEPATITIS ASSOCIATED ANTIGEN OR ANTIBODY IN A BUFFER MIXTURE;

B. COATING A TEST APPARATUS BY CONTACTING SAID TEST APPARATUS WTIH SAID SOLUTION AND C. INCUBATING SAID TEST APPARATUS WHILE IN CONTACT WITH SAID SOLUTION D. WASHING SAID CUBATED COATED TEST APPARATUS WITH A BUFFER MIXTURE;

E. PLACING SAID UNKNOWN SAMPLE IN CONTACT WITH SAID INCUBATED AND WASHED TEST APPARATUS;

F. INCUBATING SAID UNKNOWN SAMPLE WHILE IN CONTACT WITH SAID COATED TEST APPARATUS FOR FROM 0.5 TO 42 HOURS TO BOND ANY OF SAID HEPATITIS ASSOCIATED ANTIGEN OR ANTIBODY PRESENT IN SAID UNKNOWN SAMPLE TO SAID INCUBATED TEST APPARATUS;

G. WASHING SAID INCUBATED TEST APPARATUS WITH A BUFFERED SOLUTION TO REMOVE SAID UNKNOWN SAMPLE LEAVING SAID BONDED HEPATITIS ASSOCIATED ANTIGEN OR ANTIBODY ON THE COATING OF SAID TESPT APPARATUS;

H. CONTACTING SAID WASHED TEST APPARATUS WITH A PURIFIED FORM OF SAID HEPATITIS ASSOCIATED ANTIGEN OR ANTIBODY LABELED WITH THE RADIOACTIVE ISOTOPE 1-125;

I. INCUBATING SAID WASHED TEST APPARATUS WHILE IN CONTACT WITH SAID PURIFIED FORM OF SAID HEPATITIS ASSOCIATED ANTIGEN OR ANTIBODY LABELED WITH A RADIOACTIVE ISOTOPE SO AS TO BOND SAID PURIFIED FORM TO SAID ANTIGEN OR ANTIBODY BONDED ON SAID TEST APPARATUS AND THEREBY PRODUCE A RADIOCATIVELY TRACTED INCUBATED COATING;

J. WASHING SAID RADIOACTIVELY TRACED INCUBATED COATING WITH A BUFFERED SOLUTION TO REMOVE ANY UNBONDED PURIFIED FORM;

K. COUNTING RADIATION EMITTED FROM SAID RADIOCATIVELY TRACTED INCUBATED COATING;

AND L. COMPARING THE NUMBER OF COUNTS FROM SAID COATING WITH THE NUMBER OF COUNTS FROM A CONTROL SAMPLE PREPARED BY STEPS (A) TO (K).

2. The method of claim 1 wherein said test apparatus and said formed solution of said antigen or antibody are incubated for a period of from 6 to 72 hours.

3. The method of claim 1 wherein the buffer mixture of said three washing steps comprises 0.02 % by weight sodium azide in a 0.01 molar solution of 2-amino-hydroxymethyl-1, 3-propanediol hydrochloride.

4. The method of claim 1 wherein said test apparatus and said purified form are incubated for from 1 to 24 hours at room temperature.