Selective capture and enrichment of proteins expressed on the cell surface

• US 7,348,416 B2
• Filed: 11/19/2004
• Issued: 03/25/2008
• Est. Priority Date: 11/21/2003
• Status: Expired due to Fees

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Ryan, Terence, McCaffery, Ian, Domon, Bruno
• Primary Examiner(s): Bragdon; Kathleen Kerr
• Assistant Examiner(s): ROOKE, AGNES BEATA
• Application Number:
  US10/992,235
• Publication Number:
  US 20060154332A1
• Time in Patent Office: 1,222 Days
• Field of Search: 514/12
• US Class Current: 530/413
• CPC Class Codes: C07K 1/14 Extraction; Separation; Purification
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748

Abstract

The present invention provides a method for enriching cell surface proteins by oxidizing glycosylated protein with periodate. The present invention further provides that the cell surface proteins are obtained from intact cells either from cell lines or from tissues. The present invention provides a method not only capable of obtaining increased quantities of the cell surface proteins, but also capable of obtaining a wide ranges of proteins that are not detectable by other means.

Claims

1. A method for obtaining cell surface proteins comprising the steps of:

a) mixing intact cell sample with a periodate forming a mixture,

b) reacting the mixture with a reagent forming a binding complex, and wherein the reagent is selected from a group consisting of hydrazide and amine,

c) isolating the proteins with a reduction reagent from the binding complex, wherein the isolated proteins in the reduction reagent are cell surface protein.

2. The method of claim 1, wherein the intact cells are either cell lines or cells isolated from tissues.

3. The method of claim 1, wherein periodate is sodium periodate.

4. The method of claim 1, wherein the binding reagent is on a resin or a solid support.

5. The method of claim 1, wherein the cell surface proteins are 50 folds enriched relative to the total cellular proteins.

6. The method of claim 1, wherein the cell surface proteins are 100 folds enriched relative to the total cellular proteins.

7. A method for obtaining cell surface proteins comprising the steps of:

a) obtaining a homogenous cell population from a biological samples, which are intact cell,

b) mixing intact cell sample with a periodate forming a mixture,

c) reacting the mixture with a reagent forming a binding complex, and wherein the reagent is selected from a group consisting of hydrazide and amine,

d) isolating the proteins with a reduction reagent from the binding complex, wherein the isolated proteins in the reduction reagent are cell surface protein.