The determination of serum transthyretin chemical modification by capillary electrophoresis combined with MALDI-TOF/MS analysis

Abstract

Abstract: Objective　To establish a combined method for analysis of serum transthyretin(TTR) chemical modification using capillary electrophoresis (CE) with matrixassisted laser desorption ionizationtimeofflight mass spectrometry (MALDI-TOF/MS) analysis. Methods　Enzymelinked immunosorbent assay (ELISA) was used to determine serum TTR in 20 senile systemic amyloidosis (SSA) patients and 20 healthy subjects. Serum TTR was separated by CE. Determination of TTR chemical modification was analyzed by MALDI-TOF/MS. Results　The levels of serum TTR was (0.25±0.04)g/L in control group and (0.26±0.03)g/L in SSA patients, and there was no significant difference between control group and SSA patients(P>0.05). The chemical modification of TTR found 4 TTR chemical modification peaks, as native-TTR ［masstocharge ratio (m/z): 13 750.86±1.48］, sul-TTR (m/z ratio: 13 830.63±2.76), cys-TTR (m/z ratio: 13 871.70±2.70) and cysgly-TTR (m/z ratio: 13 919.00±14.77) by MALDI-TOF/MS analysis. Conclusions　The combined method of CE with MALDI-TOF/MS analysis can be used directly to determine the chemical modification of TTR.

Key words: Transthyretin, Capillary electrophoresis, Mass spectrometry, Chemical modification

ZHANG Wenliang;SONG Lin;LI Youxin;WEI Dianjun;HU Kesheng;SUN Xuguo. The determination of serum transthyretin chemical modification by capillary electrophoresis combined with MALDI-TOF/MS analysis[J]. , 2012, 27(3): 185-188.

References

［1］Sun X, Ando Y, Haraoka K, et al.　Role of VLDL/chylomicron in amyloid formation in familial amyloidotic polyneuropathy［J］. Biochem Biophys Res Commun, 2003,311(2):344-350.

［2］罗　云.联合检测血清总胆汁酸和前清蛋白在肝脏疾病诊断中的应用［J］.重庆医学，2007，36（12）：1167-1168.

［3］Ueda M, Ando Y, Haraoka K, et al. Aging and transthyretin-related amyloidosis: pathologic examinations in pulmonary amyloidosis［J］. Amyloid, 2006,13(1):24-30.

［4］Munar-Ques M, Martinez-Nadal J.Finding of vascular amyloid TTR in inferior nasal concha in a patient with FAP TTRVal30Met［J］. Amyloid, 2008,15(4):272-274. 

［5］孙续国，刘金平，宋剑蝉，等.利用MALDI-TOF/MS法测定转甲状腺素蛋白的基因突变和化学修饰［J］.中华临床防治医学杂志，2010，2(1):4-7. 

［6］Ueda M, Misumi Y, Mizuguchi M, et al. SELDI-TOF mass spectrometry evaluation of variant transthyretins for diagnosis and pathogenesis of familial amyloidotic polyneuropathy［J］. Clin Chem, 2009，55(6):1223-1227.

［7］王跃国，王惠民，张　芹，等.毛细管等速电泳法分析肾病综合征患者的脂质代谢［J］.检验医学，2009,24(1):25-28.〓

［8］赵　辉，刘金平，郑　芳，等. 家族性淀粉样多发性神经性损害转甲状腺蛋白的化学修饰［J］. 第四军医大学学报，2008,29（10）:883-885.

［9］Tsuzuki T, Mita S, Maeda S, et al. Structure of the human prealbumin gene［J］. J Biol Chem, 1985，260(22): 12224-12227.

［10］Jiang X, Smith CS, Petrassi HM, et al.An engineered transthyretin monomer that is nonamyloidogenic, unless it is partially denatured［J］. Biochemistry,2001，40(38):11442-11452.

［11］Tanskanen M, Peuralinna T, Polvikoski T, et al. Senile systemic amyloidosis affects 25% of the very aged and associates with genetic variation in alpha2-macroglobulin and tau: a population-based autopsy study［J］. Ann Med, 2008，40(3):232-239.

［12］Liu L, Wang J, Liu B, et al.Serum levels of variants of transthyretin down-regulation in cholangiocarcinoma［J］. J Cell Biochem, 2008，104(3):745-755.

［13］Lim A, Prokaeva T, McComb ME, et al. Identification of S-sulfonation and S-thiolation of a novel transthyretin Phe33Cys variant from a patient diagnosed with familial transthyretin amyloidosis［J］. Protein Sci, 2003，12(8):1775-1785.

［14］Zhang Q, Kelly JW. Cys10 mixed disulfides make transthyretin more amyloidogenic under mildly acidic conditions［J］. Biochemistry，2003, 42(29): 8756-8761.

［15］Nakanishi T, Yoshioka M, Moriuchi K, et al. S-sulfonation of transthyretin is an important trigger step in the formation of transthyretin-related amyloid fibril［J］. Biochim Biophys Acta, 2010，1804(7):1449-1456.

[1]	LI Xijiu, ZHOU Jianwei, ZHANG Qin, LONG Yunlin, WU Yingsong. Establishment and performance evaluation of UPLC-MS/MS for 6 markers in dried blood spots [J]. Laboratory Medicine, 2023, 38(5): 446-453.
[2]	XU Weixin, CHEN Qing, DU Wenyong, GAO Yan. Plasma metabolic markers in patients with insomnia disorder [J]. Laboratory Medicine, 2023, 38(5): 454-459.
[3]	WU Jie, LI Zenghe, CHEN Baorong, KONG Xujing. Establishment of traceable steroid hormone standard curve for dried blood spot samples [J]. Laboratory Medicine, 2023, 38(5): 489-493.
[4]	Expert Group on the Consensus on Androgen Testing in Polycystic Ovary Syndrome China Association for Promotion of Health Science and Technology-Fertility Protection and Preservation Committee. Consensus on androgen testing in polycystic ovary syndrome [J]. Laboratory Medicine, 2023, 38(3): 203-208.
[5]	YANG Xiaodong, LI Quanle, PAN Qingqing, ZHOU Jing, XIE Yangmin, ZOU Jihua, SHEN Min, ZHANG Man. Establishment and performance evaluation of candidate reference measurement procedure for the determination of immunosuppressive drugs in human whole blood by ID-LC-MS/MS [J]. Laboratory Medicine, 2023, 38(3): 215-222.
[6]	WANG Yajie, ZU Bailing, GAN Chenxin, WU Jiaoxiang, SHU Jie, HANG Chen, CHEN Minghui, LIN Fujun, HU Zhigang, HUANG Biao, SHENG Huiming. Identification of plasma differential peptids in PLA2R1-related membranous nephropathy patients [J]. Laboratory Medicine, 2023, 38(3): 223-229.
[7]	HUANG Yuan, CHEN Feng, MAO Leili, ZHANG Linglin, LÜ Qian, YAN Jun, CUI Wei. Application of serum peptidomics based on MALDI-TOF MS in the differential diagnosis of benign and malignant pancreatic diseases [J]. Laboratory Medicine, 2023, 38(1): 1-7.
[8]	TANG Ningbo, LIU Guancai, LIU Xiaoyu, YANG Fengfan, GONG Zhimei, SUN Keqi. Establishment and evaluation for the determination of serum digitoxin as a candidate reference method [J]. Laboratory Medicine, 2022, 37(5): 456-462.
[9]	LU Youli, YANG Shuangshuang, ZHANG Meiwei, OU Meixian, DONG Chunxia, SHEN Weiwei, JIANG Fengli, LI Shuijun. Simultaneous determination of 5 steroid hormones in human serum by ultra-performance liquid chromatography-tandem mass spectrometry and establishment of reference intervals in healthy adults [J]. Laboratory Medicine, 2022, 37(2): 165-173.
[10]	XU Runhao, ZOU Chen, CAO Yun, CHEN Feng, LIU Yijing, ZHANG Shulin, ZHANG Jie. Roles of serum amino acid determinations in the diagnosis of early non-small-cell lung cancer [J]. Laboratory Medicine, 2022, 37(12): 1157-1163.
[11]	LIU Congcong, LI Jiaping, ZHOU Hongwei. Performance evaluation of 2 mass spectrometry systems in clinical microbial identification [J]. Laboratory Medicine, 2022, 37(11): 1084-1088.
[12]	YANG Xi, SONG Wenqi, DONG Fang, MENG Qingying, ZHEN Jinghui, ZHOU Wei. Application of Gram staining and MALDI-TOF MS in the rapid identification of pathogen kinds in children's positive blood culture [J]. Laboratory Medicine, 2022, 37(10): 969-973.
[13]	YU Jiajia, LI Yuanrui, LIU Ying. MALDI-TOF MS combined with SDS and SAP pretreatment for rapid identification of positive blood culture [J]. Laboratory Medicine, 2021, 36(8): 864-868.
[14]	DING Yiwei, HAN Zhihai. Identification of MALDI-TOF MS in determining the bacteria from artificial joint replacement infection [J]. Laboratory Medicine, 2021, 36(6): 656-661.
[15]	HOU Weiwei, JIANG Lian, LI Dong. Clinical application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry in the direct detection of blood culture positive specimens [J]. Laboratory Medicine, 2021, 36(4): 424-429.