Derivatized glass supports for peptide and protein sequencing

• US 5,137,765 A
• Filed: 09/13/1990
• Issued: 08/11/1992
• Est. Priority Date: 08/05/1988
• Status: Expired due to Fees

• Current Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Original Assignee: Porton Instruments Incorporated
• Inventors: Farnsworth, Vincent
• Primary Examiner(s): CANNON, JAMES
• Application Number:
  US07/581,936
• Time in Patent Office: 698 Days
• Field of Search: 428/429, 428/64, 428/288, 428/289, 435/6, 436/89, 436/178
• US Class Current: 428/64.1
• CPC Class Codes: C07K 1/128 sequencing
• CPC Class Codes: G01N 33/552 Glass or silica
• CPC Class Codes: Y10T 428/21 Circular sheet or circular blank
• CPC Class Codes: Y10T 428/31612 As silicone, silane or siloxane
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

Peptides or proteins are sequenced by stepwise degradation while immobilized on a glass support derivatized with a silica-binding substance bearing a free acid group, especially a sulfonic acid group. The support is preferably derivatized with 2-(4-chlorosulfonyl phenyl) ethyl trimethoxysilane.

Peptide sequencing performance is improved if the support is also derivatized with a monomeric silica-binding substance bearing a free quaternary ammonium group, such as N-trimethyoxysilyl propyl -N,N,N-trimethyl ammonium chloride.

Claims

1. A derivatized glass support comprising a silicaceous support derivatized with a first silica-binding substance having a free acid group, said acid group having a pKa<

1, and a second silica-binding substance having a free quaternary ammonium group, said support being capable of retaining a peptide or protein.

2. The support of claim 1 wherein said acid group is selected from the group consisting of phosphoric, picric, hydrochloric, trifluoroacetic, trichloracetic, chromic, hydroiodic, pyrophosphoric, sulfonic and halosulfonic acids.

3. The support of claim 1 in which the acid group is a sulfonic or halosulfonic acid.

4. The support of claim 3 in which the support was prepared by incubating a glass support with a 4-chlorosulfonyl phenyl alkyl alkoxysilane.

5. The support of claim 4 in which the silane was 2-(4-chlorosulfonyl phenyl) ethyl trimethyoxysilane.

6. The support of claim 4 in which the silane was 3-(trimethylsilyl)-1-propane sulfonic acid.

7. The support of claim 1 in which the first substance is 2-(4-chlorosulfonyl phenl) ethyl trimethoxysilane, and the second substance is N-trimethoxysilyl propyl -N, N, N-trimethyl ammonium chloride.

8. The support of claim 1 in which the first or second substance was an organosilane.

9. The support of claim 8 in which the second substance was N-trimethoxysilyl propyl-N,N,N-trimethyl ammonium chloride.