Trypsin Interferon

Calf kidneys, dog kidneys and rhesus monkey kidneys were treated with trypsin to give suspensions of cells. The suspensions were centrifuged and the packed cells diluted with 400 volumes (calf cells) or 200 volumes (dog cells and rhesus monkey cells) of a growth medium consisting of 5% horse serum and 0.5% lactalbumen hydrolysate in Earle s saline, with 100 units/ml each of penicillin and streptomycin. These media were used separately to produce Semliki Forest/calf interferon, Semliki Forest/dog interferon and Semliki Forest/rhesus monkey interferon. The cell-containing growth medium was dispensed into 500 ml medical flat bottles (70 ml in each). The cultures were incubated at 36°C. Confluent sheets of cells (monolayers) were formed in 5 to 6 days. The growth medium was then removed and the monolayers were washed with isotonic phosphate-buffered saline, pH 7.5. 

Figure 1. Electropherogram of Tryptic Digest Derived from Interferon. About 1 mg/ml of recombinant interferon (Hoffmann-La Roche, Inc.), was submitted to proteolytic digestion using 40 pi of a 1 mg/ml trypsin solution in 0.05 M Tris-acetate buffer, pH 7.5. The...

Fig. 4.2 Degradation of (- -) PEGylated interferon a2b (PEG2.40 K"IFN-a2b) and (-0-) non-PEGylated interferon a2b (IFN-a2b) in the presence of trypsin at 37° C. Figure adapted from Ramon et al. (2005)...

Raehs SC, Sandow J, Wirth K, Merkle HP (1988) The adjuvant effect of bacitracin on nasal absorption of gonadorelin and buserelin in rats. Pharm Res 5 689-693 Ramon J, Saez V, Baez R, Aldana R, Hardy E (2005) PEGylated Interferon-alpha2b A Branched 40K Polyethylene Glycol Derivative. Pharm Res 22 1375-1387 Reseland JE, Holm H, Jacobsen MB, Jenssen TG, Hanssen LE (1996) Proteinase inhibitors induce selective stimulation of human trypsin and chymotrypsin secretion. J Nutr 126 634-642... 

Various enzymes, lysozyme, catalase, phosphatase, malate/formate dehydrogenase, adenylase kinase, pymvate decarboxylase, alkaline phospatase, serum albumin, lipoprotein, interferon, growth factor, polypeptide hormone DNA, RNA, y-globulin, proteinase, carboxypeptidase, endotoxins, pyrogen, IgG, trypsin inhibitor, transferrin, casein, L-benzoyl arginine ethyl ester, cytocrome C IgG, heparin... 

Inbibin (activin), beta A Inhibin (activin), beta B Inbibin (activin), beta C Inhibin (activin), beta E Inhibin, alpha Insulin C-peptide Insulin, A chain Insulin, B chain Insulin-like growth factor lA Insulin-like growth factor II Inter-alpha trypsin inhibitor, HI Inter-alpha trypsin inhibitor, H2 Inter-alpha trypsin inhibitor, H4 Inter-alpha trypsin inhibitor, L Interferon alpha Interferon beta Interferon gamma Interleukin-1 beta Interleukin-10 Interleukin-12, alpha Interleukin-12, beta Interleukin-1 receptor antagonist Interleukin-2 Interleukin-4... 

Antivirin, an unidentified material isolated from both established and primary cell lines grown in the absence of interferon-inducing factors is said to inhibit both RNA and DNA viruses [136]. The material is stable to trypsin, periodate oxidation and to heating for 1 hour at 100°C. It is a little odd that this material comes from cells which presumably will support viral growth, but the original paper is in Japanese and the abstract is uninformative. 

Affinity chromatography of Clostridium collagenase and clostripapain Purification of mouse interferon Purification by affinity chromatography of retinol-binding protein Isolation of a proviral coding strand of avian myeloblastosis virus from leukaemic chicken myeloblast Purification by affinity chromatography of trypsin... 

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