HIGH PERFORMANCE LIQUID CHROMATOGRAPHY / ELECTROSPRAY IONIZATON MASS SPECTROMETRIC CHARACTERIZATION OF RECOMBINANT L-ASPARAGINASE II
- VernacularTitle:重组L-天门冬酰胺酶II的液相色谱/电喷雾离子化质谱法分析
- Author:
Jun HAN
;
Longsheng SHENG
;
Zhongyuan YANG
;
Bingren XIANG
;
Dengkui AN
- Publication Type:Journal Article
- From:
Acta Pharmaceutica Sinica
2001;36(1):46-50
- CountryChina
- Language:Chinese
-
Abstract:
AIM To characterize the primary structure of recombinant L-asparaginase II product. METHODS The molecular weight of the protein was measured by pneumatically-assisted electrospray ionization mass spectrometry with flow injection mode. Subsequently, tryptic peptide mapping was performed by high performance liquid chromatography on a C8 column with tandem UV and MS detection. An easy-to-use and simple denaturation process with trichloroacetic acid was conducted prior to tryptic digest so as to release the digest resistance from the protein structure. The amino acid sequences of the tryptic peptides were elucidated based on their in-source collision-induced dissociation spectra. RESULTS The measured molecular mass was different from the theoretical value. Three amino acid variations were unambiguously detected along the peptide backbone derived from the gene-encoding sequence. CONCLUSION This paper revealed that LC/ESI/MS had provided a promising and robust technique in primary structure analysis and quality control of DNA-derived recombinant protein pharmaceuticals.
