Identification of protein components and quantitative immunoassay for SEC2 in staphylococcin injection.
In China, staphylococcin injection has been commonly used in combined cancer therapy to enhance the systemic immune response and reduce the toxicities associated with chemotherapy or radiation therapy in the last decade. It is claimed that the main effective component is staphylococcal enterotoxin C2 (SEC2). However, no standard method based on the concentration of SEC2 has been established for quality control of the injection products. In this study, a sensitive and reliable biotin-streptavidin-ELISA (BS-ELISA) method was established for detection and quantification of SEC2. In addition, 1-D SDS-PAGE coupled with nano-LC-MS/MS was performed to identify the protein components in the injection products from one manufacturing company. The results of the BS-ELISA showed that SEC2 only accounted for less than 0.1% of the total protein in the injection products, and the nano-LC-MS/MS results showed that fifty-five proteins of Staphylococcus aureus were confidently identified in the injection solution. Seventeen out of these proteins, including SEC2, were well-known virulence factors. In addition, eighteen proteins of other Gram-positive bacteria were also confidently identified. Thus, the results indicated that SEC2 is of very low concentration in the injection products and the process of the injection preparation should be improved for health and safety consideration.
Ding
Ding
D
Institute of Pharmacology & Toxicology and Biochemical Pharmaceutics, Zhejiang University, Hangzhou, PR China.
Huang
Peng
P
Sun
Hongying
H
Xue
Qiao
Q
Pan
Yingqiu
Y
Chen
Shuqing
S
D000818:Animals; D001430:Bacteriocins / Q000008:administration & dosage*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D036103:Nanotechnology; D053719:Tandem Mass Spectrometry
D001430:Bacteriocins; C017724:staphylococcin