Journal of Shandong University (Health Sciences) ›› 2018, Vol. 56 ›› Issue (5): 58-63.doi: 10.6040/j.issn.1671-7554.0.2017.843

Previous Articles    

Human neurofilament medium polypeptide immunogenicity and specificity of antibodies

LI Xin1, LIU Jianwei1, XUE Hao2, ZHENG Wenhui1, ZHENG Wen1, QIN Zeming1, WEN Hongling1, ZHOU Xiaoying3, WANG Zhiyu1, ZHAO Li1   

  1. 1. Department of Laboratory Science of Sanitary Microbiology, School of Public Health, Shandong University, Jinan 250012, Shandong, China;
    2. Department of Neurosurgery, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
    3. Department of Ultrasound, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2017-09-05 Published:2022-09-27

PDF (PC)

12

Abstract: Objective To explore the immunogenicity of human middle subunit of neurofilament(NFM)polypeptide and the specificity of the antibodies in order to provide basis for the preparation of monoclonal antibody(McAb)and establishment of an ELISA detection method. Methods The amino acid sequences of NFM in humans, rats and mice were analyzed. Four polypeptides(P1, P2, P3, P4)were selected, synthesized and coupled with KLH. Then the coupled polypeptides were used as antigens to immunize mice. After immunization, the mice serum was harvested, and the titer of the antibodies were determined with ELISA. The reactivity of antibodies to humans, rats, mice and NFM expressed in vitro were detected with Western blotting and immunohistochemistry(IHC). Results The four peptides had 山 东 大 学 学 报 (医 学 版)56卷5期 -李昕,等.人神经丝中分子量亚单位多肽的免疫原性及其抗体的特异性 \=-strong immunogenicity, and antibodies of high titer were obtained. The IHC showed that all of the four antibodies were able to respond to NFM in human glioma, rat and mouse brain tissues. Western blotting indicated that anti-P1 and anti-P4 antibodies could react with the NFM in human glioma, rat and mouse brain tissues, and the anti-P1 antibody had two strips. Anti-P1, anti-P2 and anti-P3 could react with NFM expressed in vitro. Conclusion The four peptides have strong immunogenicity. The antibodies produced by mice can respond to NFM in human glioma, rat and mouse brain tissues and NFM expressed in vitro. They can be used in preparing McAb.

Key words: Middle subunit of neurofilament, Pepitide, Immunogenicity, Antibody specificity

CLC Number: 

  • R392-33
[1] Yuan A, Rao MV, Veeranna, et al. Neurofilaments at a glance[J]. J Cell Sci, 2012, 125(14): 3257-3263.
[2] Rao MV, Yuan AD, Campbell J, et al. The C-Terminal domains of NF-H and NF-M subunits maintain axonal neurofilament content by blocking turnover of the stationary neurofilament network[J]. PLoS One, 2012, 7(9): 10.
[3] Perrot R, Berges R, Bocquet A, et al. Review of the multiple aspects of neurofilament functions, and their possible contribution to neurodegeneration[J]. Mol Neurobiol, 2008, 38(1): 27-65.
[4] Yuan AD, Rao MV, Sasaki T, et al. Alpha-Internexin is structurally and functionally associated with the neurofilament triplet proteins in the mature CNS[J]. J Neurosci, 2006, 26(39): 10006-10019.
[5] Yuan AD, Sasaki T, Kumar A, et al. Peripherin is a subunit of peripheral nerve Neurofilaments: implications for differential vulnerability of CNS and peripheral nervous system axons[J]. J Neurosci, 2012, 32(25): 8501-8508.
[6] 陈建国, 滕俊琳, 翟中和, 等. 神经丝蛋白NF-M的功能分析[J]. 科学通报, 1996, 41(3): 263-266.
[7] Laser-Azogui A, Kornreich M, Malka-Gibor E, et al. Neurofilament assembly and function during neuronal development[J]. Curr Opin Cell Biol, 2015, 32: 92-101.
[8] Ludemann N, Clement A, Hans VH, et al. O-glycosylation of the tail domain of neurofilament protein M in human neurons and in spinal cord tissue of a rat model of amyotrophic lateral sclerosis(ALS)[J]. J Biol Chem, 2005, 280(36): 31648-31658.
[9] Deng YQ, Li B, Liu F, et al. Regulation between O-GlcNAcylation and phosphorylation of neurofilament-M and their dysregulation in Alzheimer disease[J]. FASEB J, 2008, 22(1): 138-145.
[10] 苏闻, 陈海波, 李淑华, 等. 神经丝蛋白与帕金森病的相关性研究[J]. 中国神经免疫学和神经病学杂志, 2010, 17(3): 177-179. SU Wen, CHEN Haibo, LI Shuhua, et al. Study on relationship between neurofilament Proteins and Parkinson disease[J]. Chin J Neuroimmunol & Neurol, 2010, 17(3):177-179.
[11] Niemelä V, Landtblom AM, Blennow K, et al. Tau or neurofilament light-Which is the more suitable biomarker for Huntingtons disease?[J]. PLoS One, 2017, 12(2): 11. doi:10.1371/journal.pone.0172762.
[12] Shahim P, Zetterberg H, Tegner Y, et al. Serum neurofilament light as a biomarker for mild traumatic brain injury in contact sports[J]. Neurology, 2017, 88(19): 1788-1794.
[13] Kuhle J, Barro C, Andreasson U, et al. Comparison of three analytical platforms for quantification of the neurofilament light chain in blood samples: ELISA, electrochemiluminescence immunoassay and Simoa[J]. Clin Chem Lab Med, 2016, 54(10): 1655-1661.
[14] Rudrabhatla P, Grant P, Jaffe H, et al. Quantitative phosphoproteomic analysis of neuronal intermediate filament proteins(NF-M/H)in Alzheimers disease by iTRAQ[J]. FASEB J, 2010, 24(11): 4396-4407.
[15] Martinez-Morillo E, Childs C, Garcia B P, et al. Neurofilament medium polypeptide(NFM)protein concentration is increased in CSF and serum samples from patients with brain injury[J]. Clin Chem Lab Med, 2015, 53(10): 1575-1584.
[16] Funes SC, Chiari ME, Comin R, et al. Experimental Guillain-Barre syndrome induced by immunization with gangliosides: keyhole limpet hemocyanin is required for disease triggering[J]. Biochim Biophys Acta, 2017, 1863(6): 1473-1478.
[17] Markl J, Lieb B, Gebauer W, et al. Marine tumor vaccine carriers: structure of the molluscan hemocyanins KLH and HtH[J]. J Cancer Res Clin Oncol, 2001,127 Suppl 2:R3-9.
[18] Harris JR, Markl J. Keyhole limpet hemocyanin(KLH): a biomedical review[J]. Micron, 1999, 30(6): 597-623.
[19] Tripathi NK. Production and purification of recombinant proteins from escherichia coli[J]. Chembioeng Reviews, 2016, 3(3): 116-133.
[20] 王昆, 梁朋娟, 李尚勇, 等. 碱性蛋白酶抑制剂lupI-MSSS原核表达载体的构建及其蛋白表达、纯化[J]. 食品与发酵工业, 2017, 43(7): 20-26.
[1] JIANG Jinzhu1， WU Yan2， LU Qing1， JIA Jun1, SHAO Yang1， SONG Guodong1. Effect of matrix metalloproteinase-7 on immunogenicity of porcine acellular dermal matrix [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(8): 74-.
[2] GU Yan-zheng1,3, CAI Yan2, NI Li2, HUANG Chen2, YANG Hui-lin2,3, ZHANG Xue-guang1,3, SHI Qin2,3. Alterations of immunogenicity in the differentiation from human bone marrow derived mesenchymal stem cells to chondrocytes [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2010, 48(11): 24-28.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd