An improved process for purifying proteins, particularly proteins having molecular weights in excess of 12,000 involves novel applications of high performance liquid chromatography on a preparative scale to provide homogeneous end product in excellent yield. In an embodiment of the process, human interferon is produced as a homogeneous protein.
RELATED APPLICATIONS
This is a continuation of application Ser. No. 247,442, now abandoned, Mar. 25, 1981, which is a divisional of Ser. No. 167,165, filed 7/9/80, now U.S. Pat. No. 4,289,690, which is a continuation-in-part of Ser. No. 106,644 filed 12/26/79, now abandoned, which is a continuation-in-part of Ser. No. 77,710 filed 9/21/79, now abandoned, which is a continuation-in-part of Ser. No. 62,374 filed 7/31/79, now abandoned, which is a continuation-in-part of Ser. No. 963,257 filed 11/24/78. now abandoned.
This invention relates to an agarase enzyme system purified from bacterial strain 2-40, which has a high level of activity for the depolymerization of complex polysaccharides, including agar and agarose. Further, the invention relates to methods of purifying, defining, characterizing and assaying the agarase enzyme system and the encoding gene(s). Finally, the invention relates to methods of using the purified agarase enzyme system.
A purified DNA molecule encoding a glycoprotease from Pasteurella haemolytica is disclosed. The DNA comprises a sequence of approximately 975 base pairs coding for a glycoprotease having a molecular weight of approximately 35.2 kD. The glycoprotease is specific for cleaving O-glycosylated carbohydrate portions from O-glycoproteins. The glycoprotease has a major cleavage site in glycophorin A between Arg31 and Asp32.
Condylomata Acuminata infections (anogenital warts) are treated in infected patients by administering liquid nitrogen and immediately thereafter beginning administering recombinant DNA human alpha interferon thrice a week for three weeks. The interferon exemplified is recombinant DNA human interferon alfa-2b in which 1.0.times.10.sup.6 International Units are administered by injection to each lesion. The liquid nitrogen is the cryosurgical agent exemplified and it is topically administered to each lesion by conventional means.
Human leukemia T-cells and B-cells are inhibited from proliferating by treatment with a combination of recombinant human alpha and gamma interferons, either simultaneously or sequentially, and the alpha interferon is preferably recombinant human alfa-2b interferon.
The gene for Streptococcus pyogenes DNase B has been cloned and vectors incorporating the cloned DNA have been used to transform Escherichia coli, allowing efficient and rapid production of the DNase in E. coli without the necessity of growing large quantities of S. pyogenes. The enzyme can be produced with a leader peptide at its amino-terminus. An improved method for the purification of naturally occurring S. pyogenes DNase B enzyme is also provided. The DNase B enzyme produced, either by purification of naturally occurring enzyme or by recombinant DNA techniques, can be used to generate antibodies and can also be used in immunochemical assays to detect the presence of anti-DNase B antibodies in serum as a marker of infection by S. pyogenes.
