Sequential oligonucleotide syntheses using immunoaffinity techniques

• US 5,464,759 A
• Filed: 02/17/1993
• Issued: 11/07/1995
• Est. Priority Date: 12/21/1988
• Status: Expired due to Fees

• Current Assignee: Bionebraska, Inc., Board of Regents of the University of Nebraska
• Original Assignee: Bionebraska, Inc., Board of Regents of the University of Nebraska
• Inventors: Wylie, Dwane, Lewis, William, Coolidge, Thomas R., Wagner, Fred W., Stout, Jay, Schuster, Sheldon M., van Heeke, Gino
• Primary Examiner(s): Parr, Margaret
• Assistant Examiner(s): Marschel, Ardin H.
• Application Number:
  US08/018,100
• Time in Patent Office: 993 Days
• Field of Search: 435/6, 435/91, 435/91.2, 536/22.1, 536/23.1, 536/24.1, 536/24.2, 536/24.31, 536/24.32, 536/25.3, 935/78, 935/88, 935/77, 514/44
• US Class Current: 435/91.2
• CPC Class Codes: C07K 1/006 of peptides containing derivatised side chain amino acids
• CPC Class Codes: C07K 1/04 on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/824 Immunological separation techniques

Abstract

The invention is directed to a method of purifying sequentially synthesized peptides and oligonucleotides by affinity techniques. Selected products are capped with and N-terminus capping agent for peptides or a 5'"'"'-terminus capping agents for oligonucleotides, and then bound with affinity agents that are selective for the corresponding capping agents.

Claims

1. A method for DNA replication comprising:

applying to the DNA to be replicated, a polymerase chain reaction technique in combination with an oligonucleotide probe produced by a method comprising;

(a) preparing a 3'"'"'-blocked nucleotide compound of at least one nucleotide in length;

(b) reacting a 3'"'"'-activated, 5'"'"'-blocked nucleotide with the 3'"'"'-blocked nucleotide compound to produce a mixture of an extended nucleotide compound and an unreacted nucleotide compound;

(c) adding a 5'"'"'-terminus capping agent to the mixture to cap the unreacted nucleotide compound;

(d) combining the mixture of the extended nucleotide compound and the 5'"'"'-capped unreacted nucleotide compound with an affinity agent that is selective for the cap on the 5'"'"'-capped. unreacted nucleotide; and

(e) isolating the extended nucleotide compound thereby producing the oligonucleotide.

2. The method of claim 1 wherein the replicated DNA is further used in a diagnostic for a genetic disorder in a human or an animal.