US 12,449,423 B2
Qualitative analysis of proteins
Lee Hoang, Santa Clara, CA (US); Chris Suh, San Jose, CA (US); and Douglas T. Gjerde, Saratoga, CA (US)
Assigned to Phynexus, Inc., San Jose, CA (US)
Appl. No. 17/047,462
Filed by Biotage AB, Uppsala (SE)
PCT Filed Apr. 29, 2019, PCT No. PCT/EP2019/060883
§ 371(c)(1), (2) Date Oct. 14, 2020,
PCT Pub. No. WO2019/211223, PCT Pub. Date Nov. 7, 2019.
Prior Publication US 2021/0148925 A1, May 20, 2021
Int. Cl. C07K 1/16 (2006.01); G01N 33/68 (2006.01); G01N 35/00 (2006.01); G01N 35/04 (2006.01)
CPC G01N 33/6848 (2013.01) [C07K 1/16 (2013.01); G01N 35/00 (2013.01); G01N 2035/0434 (2013.01); G01N 2035/0436 (2013.01)] 15 Claims
 
1. A method for qualitative analysis of a sample protein, which method comprises the steps of
a) Providing a water swollen gel column bed comprising at least one bound protease;
b) Providing at least one sample protein in liquid buffer;
c) Digesting said sample protein by contact with the gel bed of step a) to provide for cleaving of the protein(s) into polypeptides; and
d) Subjecting the polypeptides obtained from step c) to mass spectrometry (MS) to obtain mass information related thereto,
wherein step c) comprises back and forth flow at a temperature of less than about 37° C.,
wherein the water swollen gel column bed is selected from cellulose, agarose, or dextran, and
wherein said at least one bound protease is selected from the group consisting of trypsin; Glu-C; Lys-N; Lys-C; Asp-N; Arg-C; and chymotrypsin.