IDENTIFICATION AND QUANTIFICATION OF METABOLIC
MODIFICATION OF SERUM PROTEIN AS A PROGNOSTIC BIOMARKER BY LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY.
HSIN CHEN, SHU-HUI CHEN*
NATIONAL CHENG KUNG UNIVERSITY, TAIWAN
Its has been found that estrogen metabolism such as Catechol estrogens (CEs) can conjugate with Cysteine, Lysine and Histidine to form the adducts through covalent bound, which are regarded as the initiators of the cancer. Since estrogen metabolism can react with human serum protein to form trace amount of estrogenized protein adducts,our lab developed an analysis method of liquid chromatography-mass spectrometry combined Solid Phase Extract(SPE) and Multiple Reaction Monitoring(MRM).
Human Serum Albumin (HAS) activated by 4-hrdroxyestradiol(4-OHE2) are selected as a standard so that its analysis data obtain from high resolution mass spectrometry compared with the data [1] we previous published to set up MRMs parameter. Additionally, SPE was used to purify the sample and minimize the Matrix Effect.
This analysis method was applied to measure the estrogenization percentage of HSA derived from 30 diabetic obese patients against those derived from 30 normal ones. The results indicated that the estrogenization percentage of the patient group was twice greater than the normal ones. Further, we use Standard Addition Method and External Standard Method to quantify the amount of the estrogenization protein in the serum.
References
[1] Fang, C. M.; Ku, M. C.; Chang, C. K.; Liang, H. C.; Wang, T. F.; Wu, C. H.; Chen, S. H., Identification of En-dogenous Site-specific Covalent Binding of Catechol Estrogens to Serum Proteins in Human Blood, Toxicological sciences : an official journal of the Society of Toxicology., 148 (2) 433–42, 2015.
(S. Chen, S. H. Chen) No.1, University Road, Tainan City 701, Taiwan (R.O.C.) E-mail address: [email protected]
Key words and phrases. Estrogenized Human Serum Albumin, Solid-Phase Extraction, Multiple Reaction Mon-itoring (MRM), Quantified of the serum protein.
