树鼩Vasorin单克隆抗体的制备及其应用探索

doi:10.12300/j.issn.1674-5817.2025.027

实验动物与比较医学 ›› 2025, Vol. 45 ›› Issue (5): 611-622.DOI: 10.12300/j.issn.1674-5817.2025.027

树鼩Vasorin单克隆抗体的制备及其应用探索

欧美珍¹, 李泳锋², 温莎¹, 廖舟翔², 黄雪静¹, 何敏¹^,²^,³^,⁴()(), 杨丽超¹^,³()()

^1.广西医科大学实验动物中心, 南宁 530021
^2.广西医科大学公共卫生学院, 南宁 530021
^3.区域性高发肿瘤早期防治研究教育部重点实验室/广西重点实验室, 南宁 530021
^4.靶向肿瘤学国家重点实验室, 南宁 530021

收稿日期:2025-03-01 修回日期:2025-04-15 出版日期:2025-10-25 发布日期:2025-10-23
通讯作者:
何敏（1966—），女，博士，教授，博士生导师，博士后合作导师，研究方向：比较医学。E-mail：hemin@gxmu.edu.cn。ORCID：0000-0002-2735-2474；

杨丽超（1986—），女，博士，副教授，硕士生导师，研究方向：人类疾病动物模型。E-mail：yanglichao@gxnu.edu.cn。ORCID：0000-0003-3096-9632
作者简介:欧美珍（1998—），女，硕士研究生，研究方向：比较医学。E-mail：1932476367@qq.com
基金资助:
国家自然科学基金项目“Vasorin与锌α2糖蛋白相互作用调控肝脏脂质代谢的机制研究”(32460220);广西重点研发项目“新型高效肝癌动物模型的制备与应用”(桂科AB24010127)

Preparation of Monoclonal Antibody to Vasorin in Tree Shrew and Exploration of Its Application

OU Meizhen¹, LI Yongfeng², WEN Sha¹, LIAO Zhouxiang², HUANG Xuejing¹, HE Min¹^,²^,³^,⁴()(), YANG Lichao¹^,³()()

^1.Laboratory Animal Center, Guangxi Medical University, Nanning 530021, China
^2.School of Public Health, Guangxi Medical University, Nanning 530021, China
^3.Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor, Ministry of Education(Guangxi Key Laboratory), Nanning 530021, China
^4.State Key Laboratory of Targeted Oncology, Nanning 530021, China

Received:2025-03-01 Revised:2025-04-15 Published:2025-10-25 Online:2025-10-23
Contact: HE Min (ORCID: 0000-0002-2735-2474), E-mail: hemin@gxmu.edu.cn;
YANG Lichao (ORCID: 0000-0003-3096-9632), E-mail: yanglichao@gxmu.edu.cn;

摘要/Abstract

摘要：

目的通过原核表达和纯化得到树鼩Vasorin（VASN）重组蛋白，以此蛋白免疫小鼠制备抗树鼩VASN单克隆抗体，并初步评估其应用价值。方法采用反转录聚合酶链式反应（reverse transcription-polymerase chain reaction，RT-PCR）法体外扩增树鼩VASN基因全长序列，将树鼩VASN基因片段插入到pET-30a载体中，构建pET-30a-VASN重组质粒，用BamHⅠ和SalⅠ对重组质粒进行双酶切鉴定，并通过测序进一步鉴定其正确性；将测序正确的重组质粒转化至BL21（DE3）感受态细胞中，利用异丙基-β-D-硫代半乳糖苷（isopropyl β-D-thiogalactoside，IPTG）诱导表达重组蛋白VASN；通过SDS-PAGE分离蛋白VASN，并利用KCl纯化重组蛋白VASN；用纯化的重组蛋白VASN对BALB/c小鼠进行4次免疫，并通过酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）检测小鼠血清抗体效价；取血清抗体效价达到1∶10 000以上的小鼠的脾脏细胞与骨髓瘤细胞进行细胞融合，先利用含黄嘌呤、氨基蝶呤及胸腺嘧啶核苷（hypoxanthine-aminopterin-thymidine，HAT）的培养基筛选杂交瘤细胞，随后通过ELISA筛选出能够分泌特异性抗体的阳性杂交瘤细胞株，并通过有限稀释法进行亚克隆，获得单克隆杂交瘤细胞株；通过腹水诱生法制备大量VASN单克隆抗体，利用rProtein G纯化抗体，采用ELISA和蛋白质印迹法检测单克隆抗体的结合常数和体外反应的特异性。结果成功扩增出树鼩VASN基因并构建了树鼩pET-30a-VASN重组质粒，重组质粒经测序得到的序列与树鼩VASN目的基因序列完全一致；重组蛋白VASN主要以包涵体的形式存在，纯化后的蛋白纯度达到90%，符合后续免疫实验的要求；利用重组蛋白VASN 4次免疫小鼠后，小鼠血清抗体效价达1∶729 000；利用杂交瘤技术及有限稀释法获得单克隆阳性杂交瘤细胞株，通过腹水诱生和纯化得到单克隆抗体，ELISA测定单克隆抗体的结合常数值达2.59×10⁷ L/mol；蛋白质印迹法结果显示，树鼩VASN单克隆抗体能与树鼩VASN重组蛋白结合，与猪视黄醇结合蛋白4重组蛋白、人源VASN-富含亮氨酸重复序列重组蛋白以及牛血清白蛋白均无结合反应；而抗树鼩VASN单克隆抗体能特异性识别树鼩心脏、肝脏、脾脏、肺脏、肾脏和肌肉组织中的VASN蛋白，且条带明显、背景清晰。免疫组织化学检测结果显示，该单克隆抗体可以识别蛋白VASN mRNA表达水平较高的树鼩脾脏、肺脏和树鼩永生化成纤维细胞中的VASN蛋白。结论成功制备了抗树鼩VASN的单克隆抗体，该抗体可用于树鼩永生化成纤维细胞、树鼩脾脏组织和肺脏组织的免疫组织化学检测，为进一步探索VASN在树鼩模型中的功能提供了重要的工具。

关键词: Vasorin, 树鼩, 单克隆抗体, 原核表达, 杂交瘤融合

Abstract:

Objective To obtain tree shrew Vasorin (VASN) recombinant protein through prokaryotic expression and purification, prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein, and preliminarily evaluate its application value. Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the full-length sequence of tree shrew VASN gene in vitro. The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid. The recombinant plasmid was subjected to double digestion with BamHⅠ and SalⅠ for identification, and its correctness was further verified by sequencing. The recombinant plasmid with correct sequencing was transformed into BL21 (DE3) competent cells, and isopropyl β-D-thiogalactoside (IPTG) was used to induce expression of VASN recombinant protein. Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the VASN recombinant protein was purified by KCl. Purified recombinant protein was used to immunize BALB/c mice for four times, and serum antibody titer was detected by enzyme-linked immunosorbent assay (ELISA). Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells. Hypoxanthine-aminopterin-thymidine (HAT) culture medium was first used to screen hybridoma cells. ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies, and monoclonal hybridoma cell lines were obtained by limiting dilution method. VASN monoclonal antibodies were prepared in large quantities by ascites induction method, purified using rProtein G, and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting. Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed. The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence. Recombinant protein VASN mainly existed in the form of inclusion bodies, and the purity after purification reached 90%, meeting the requirements of subsequent immunization experiments. After four immunizations with recombinant protein VASN, mouse serum antibody titer reached 1:729 000. Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification, and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x10⁷ L/mol. Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein, but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein, human VASN-leucine rich repeat recombinant protein, or bovine serum albumin. Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart, liver, spleen, lung, kidney and muscle, with clear bands and clean background. Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen, lung, and tree shrew immortalized fibroblasts with high VASN mRNA expression levels, and the detection results were positive. Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared. This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts, spleen tissue, and lung tissue, providing an important tool for further research on the function of VASN in tree shrew models.

Key words: Vasorin, Tree shrew, Monoclonal antibody, Prokaryotic expression, Hybridoma fusion

中图分类号:

Q95-33

欧美珍,李泳锋,温莎,等. 树鼩Vasorin单克隆抗体的制备及其应用探索[J]. 实验动物与比较医学, 2025, 45(5): 611-622. DOI: 10.12300/j.issn.1674-5817.2025.027.

OU Meizhen,LI Yongfeng,WEN Sha,et al. Preparation of Monoclonal Antibody to Vasorin in Tree Shrew and Exploration of Its Application[J]. Laboratory Animal and Comparative Medicine, 2025, 45(5): 611-622. DOI: 10.12300/j.issn.1674-5817.2025.027.

图/表 4

图1 pET-30a-VASN质粒的构建注：A，VASN基因扩增产物（M，DNA分子量标准；1，VASN扩增产物）；B，单菌落聚合酶链式反应鉴定结果（1~5泳道为不同菌落）；C，重组质粒双酶切鉴定结果（1~2泳道为pET-30a-VASN双酶切产物）。

Figure 1 Construction of the pET-30a-VASN recombinant plasmidNote： A， VASN gene amplification product （M， DNA Marker； 1， VASN amplification product）； B， PCR identification of monoclonal colonies （1-5， different colonies）； C， Recombinant plasmid double digestion identification （1-2， pET-30a-VASN double digestion product）.

图2 VASN重组蛋白的表达与纯化注：A，VASN重组蛋白的表达（M，蛋白质分子量标准；1，未诱导的pET-30a-VASN上清；2，未诱导的pET-30a-VASN沉淀；3，经诱导的pET-30a-VASN上清；4，经诱导的pET-30a-VASN沉淀）；B，VASN重组蛋白的纯化（M，蛋白质分子量标准；1，VASN重组蛋白纯化前；2，VASN重组蛋白纯化后；3，蛋白上样缓冲液）。

Figure 2 Expression and purification of VASN recombinant proteinNote： A， Expression of VASN recombinant protein （M， Protein marker； 1， Uninduced pET-30a-VASN supernatant； 2， Uninduced pET-30a-VASN precipitate； 3， Induced pET-30a-VASN superna-tant； 4， Induced pET-30a-VASN precipitate）； B， Purification of VASN recombinant protein （M， Protein marker； 1， Before purifi-cation of VASN recombinant protein； 2， After purification of VASN recombinant protein； 3， Protein loading buffer）.

图3 树鼩VASN单克隆抗体的制备注：A，小鼠血清效价；B，小鼠脾脏细胞和SP2/0骨髓瘤细胞融合过程；C，单克隆抗体腹水纯化（M，蛋白质分子量标准，1为纯化TV5-B11）；D，单克隆抗体结合常数测定；E，抗体特异性鉴定（M，蛋白质分子量标准；1，树鼩VASN重组蛋白；2，猪RBP4重组蛋白；3，人VASN-LRR重组蛋白；4，BSA标准蛋白）；F，抗体梯度浓度鉴定（M，蛋白质分子量标准；1，0.5 μg；2，1.0 μg；3，1.5 μg；4，2.0 μg）。

Figure 3 Preparation of monoclonal antibody to tree shrews VASNNote： A， Mouse serum titer； B， Fusion process of mouse spleen cells and SP2/0 myeloma cells； C， Monoclonal antibody ascites purified （M， Protein marker； 1， Purified TV5-B11）； D， Monoclonal antibody affinity measurement； E， Antibody specificity identification （M， Protein marker； 1， tree shrews VASN recombinant protein； 2， porcine retinol-binding protein 4 recombinant protein； 3， human VASN-leucine rich repeat recombinant protein； 4 bovine serum albumin standard protein）； F， Antibody gradient concentration identification （M， Protein marker； 1， 0.5 μg； 2， 1.0 μg； 3， 1.5 μg； 4， 2.0 μg）.

图4 树鼩VASN抗体的应用注：A，蛋白质印迹法鉴定树鼩各组织中VASN的蛋白表达水平（M，蛋白质分子量标准；1，心脏组织；2，肝脏组织；3，脾脏组织；4，肺脏组织；5，肾脏组织；6，肌肉组织）；B，实时荧光定量PCR检测树鼩各组织中VASN mRNA的相对表达水平（与肺脏组织比较，***P<0.001，****P<0.000 1；与脾脏组织比较，###P<0.001，####P<0.000 1）；C，VASN基因扩增结果（M，DNA分子量标准；1，阳性对照；2，VASN扩增产物；3，空白对照）；D，抗体应用于免疫组织化学鉴定树鼩永生化成纤维细胞中VASN蛋白的表达；E，抗体应用于免疫组织化学鉴定脾脏组织中VASN蛋白的表达；F，抗体应用于免疫组织化学鉴定肺脏组织中VASN蛋白的表达。

Figure 4 Application of tree shrews VASN antibodyNote：A， The protein expression level of VASN in various tissues of tree shrew was identified by Western blotting（M， Protein marker； 1， Heart tissue； 2， Liver tissue； 3， Spleen tissue； 4， Lung tissue； 5， Kidney tissue； 6， Muscle tissue）； B， Relative expression levels of VASN mRNA in various tissues of tree shrews detected by qRT-PCR （Compared with lung tissue，***P<0.001，****P<0.000 1； compared with spleen tissue， ###P<0.001，####P<0.000 1）； C， Amplification results of the VASN gene（M， DNA Marker； 1， Positive control； 2， VASN amplification product； 3， Blank control）； D， Antibodies were used in immunohistochemistry to identify the expression of VASN protein in immortalized fibroblasts of tree shrews； E， Antibodies were used in immunohistochemistry to identify the expression of VASN protein in spleen tissue； F， Antibodies were used in immunohistochemistry to identify the expression of VASN protein in lung tissue.

参考文献 35

[1]	黄晓燕, 徐娟, 孙晓梅, 等. 树鼩在人类疾病动物模型中应用研究进展[J]. 实验动物科学, 2013, 30(2):59-64. DOI: 10.3969/j.issn.1006-6179.2013.02.014 .
	HUANG X Y, XU J, SUN X M, et al. Development of application of tree shrew in human disease animal models research[J]. Lab Anim Sci, 2013, 30(2):59-64. DOI: 10.3969/j.issn.1006-6179.2013.02.014 .
[2]	王文广, 匡德宣, 仝品芬, 等. 树鼩的标准化研究与应用进展[J]. 实验动物科学, 2020, 37(1):74-78. DOI: 10.3969/j.issn.1006-6179.2020.01.016 .
	WANG W G, KUANG D X, TONG P F, et al. Standardization and application progress of tree shrew[J]. Lab Anim Sci, 2020, 37(1):74-78. DOI: 10.3969/j.issn.1006-6179.2020.01.016 .
[3]	苏傲蕾, 秦银鸽, 郑禹, 等. 树鼩的生物学特性研究概述[J]. 动物医学进展, 2014, 35(10):115-118. DOI: 10.16437/j.cnki.1007-5038.2014.10.031 .
	SU A L, QIN Y G, ZHENG Y, et al. Introduction to biological characteristics of tree shrew[J]. Prog Vet Med, 2014, 35(10):115-118. DOI: 10.16437/j.cnki.1007-5038.2014.10.031 .
[4]	HEIDARI F, MADADI S, ALIZADEH N, et al. The potential of monoclonal antibodies for colorectal cancer therapy[J]. Med Oncol, 2023, 40(9):273. DOI:10.1007/s12032-023-02151-1 .
[5]	黄建洪, 张春阳. 单克隆抗体在肾移植免疫抑制治疗中的应用进展[J]. 医学综述, 2008, 14(5):752-754.
	HUANG J H, ZHANG C Y. The application of monoclonal antibodies in immunosuppressive strategies of renal transplantation[J]. Med Recapitul, 2008, 14(5):752-754.
[6]	PAUL S, KONIG M F, PARDOLL D M, et al. Cancer therapy with antibodies[J]. Nat Rev Cancer, 2024, 24(6):399-426. DOI:10.1038/s41568-024-00690-x .
[7]	SORBARA M, CORDELIER P, BERY N. Antibody-based approaches to target pancreatic tumours[J]. Antibodies, 2022, 11(3):47. DOI: 10.3390/antib11030047 .
[8]	KLEIN C, BRINKMANN U, REICHERT J M, et al. The present and future of bispecific antibodies for cancer therapy[J]. Nat Rev Drug Discov, 2024, 23(4):301-319. DOI: 10.1038/s41573-024-00896-6 .
[9]	GUO X H, WU Y, XUE Y, et al. Revolutionizing cancer immunotherapy: unleashing the potential of bispecific anti-bodies for targeted treatment[J]. Front Immunol, 2023, 14:1291836. DOI: 10.3389/fimmu.2023.1291836 .
[10]	曹颖颖, 李宝莹, 王婷, 等. 瑶山亚种树鼩IFN-β和IFN-γ原核表达及其多克隆抗体制备[J]. 南方农业学报, 2022, 53(6):1713-1723. DOI: 10.3969/j.issn.2095-1191.2022.06.025 .
	CAO Y Y, LI B Y, WANG T, et al. Prokaryotic expression and polyclonal antibody preparation of IFN-β and IFN-γ of Tupaia belangeri yaoshanensis[J]. J South Agric, 2022, 53(6):1713-1723. DOI: 10.3969/j.issn.2095-1191.2022.06.025 .
[11]	曹颖颖, 李慧君, 李宝莹, 等. 瑶山亚种树鼩ISG15蛋白表达及其多克隆抗体制备[J]. 中国畜牧兽医, 2022, 49(1):273-282. DOI: 10.16431/j.cnki.1671-7236.2022.01.030 .
	CAO Y Y, LI H J, LI B Y, et al. Expression of ISG15 protein of Tupaia belangeri yaoshanensis and preparation of its polyclonal antibody[J]. China Anim Husb Vet Med, 2022, 49(1):273-282. DOI: 10.16431/j.cnki.1671-7236.2022.01.030 .
[12]	吴晋元, 周艳, 解裕萍, 等. 树鼩IgG纯化鉴定及其多克隆抗体制备和检测[J]. 现代生物医学进展, 2016, 16(7):1201-1204. DOI: 10.13241/j.cnki.pmb.2016.07.001 .
	WU J Y, ZHOU Y, XIE Y P, et al. Purification of tree shrews(Tupaia) immunoglobulin G and preparation of anti-IgG polyclonal antibody[J]. Prog Mod Biomed, 2016, 16(7):1201-1204. DOI: 10.13241/j.cnki.pmb.2016.07.001 .
[13]	MALAPEIRA J, ESSELENS C, BECH-SERRA J J, et al. ADAM17 (TACE) regulates TGF-β signaling through the cleavage of vasorin[J]. Oncogene, 2011, 30(16):1912-1922. DOI: 10.1038/onc.2010.565 .
[14]	IKEDA Y, IMAI Y, KUMAGAI H, et al. Vasorin, a transforming growth factor beta-binding protein expressed in vascular smooth muscle cells, modulates the arterial response to injury in vivo [J]. Proc Natl Acad Sci USA, 2004, 101(29):10732-10737. DOI:10.1073/pnas.0404117101 .
[15]	QIN Z X, ZHONG Y, LI P W, et al. Vasorin promotes endothelial differentiation of glioma stem cells via stimulating the transcription of VEGFR2[J]. FASEB J, 2024, 38(10): e23682. DOI: 10.1096/fj.202400159R .
[16]	LIANG W Y, ZUO J, LIU M K, et al. VASN promotes colorectal cancer progression by activating the YAP/TAZ and AKT signaling pathways via YAP[J]. FASEB J, 2023, 37(1): e22688. DOI: 10.1096/fj.202201181R .
[17]	CHEN W S, WANG Q, XU X L, et al. Vasorin/ATIA promotes cigarette smoke-induced transformation of human bronchial epithelial cells by suppressing autophagy-mediated apop-tosis[J]. Transl Oncol, 2020, 13(1):32-41. DOI: 10.1016/j.tranon.2019.09.001 .
[18]	YANG L C, CHENG X J, SHI W, et al. Vasorin deletion in C57BL/6J mice induces hepatocyte autophagy through glycogen-mediated mTOR regulation[J]. Nutrients, 2022, 14(17):3600. DOI: 10.3390/nu14173600 .
[19]	WAN F J, LI H, HUANG S P, et al. Vasorin promotes proliferation and migration via STAT3 signaling and acts as a promising therapeutic target of hepatocellular carcinoma[J]. Cell Signal, 2023, 110:110809. DOI: 10.1016/j.cellsig.2023.110809 .
[20]	WU D N, ZHANG K L, CHEN R H, et al. VASN promotes the aggressive phenotype in ARID1A-deficient lung adenocar-cinoma[J]. BMC Cancer, 2024, 24(1):1327. DOI: 10.1186/s12885-024-13083-y .
[21]	GUO X P, SUN J M, LIANG J N, et al. Vasorin contributes to lung injury via FABP4-mediated inflammation[J]. Mol Biol Rep, 2022, 49(10):9335-9344. DOI: 10.1007/s11033-022-07780-9 .
[22]	YAO Y G, LU L, NI R J, et al. Study of tree shrew biology and models: a booming and prosperous field for biomedical research[J]. Zool Res, 2024, 45(4):877-909. DOI: 10.24272/j.issn.2095-8137.2024.199 .
[23]	TONG Y H, HAO J J, TU Q, et al. A tree shrew glioblastoma model recapitulates features of human glioblastoma[J]. Oncotarget, 2017, 8(11):17897-17907. DOI: 10.18632/oncotarget.15225 .
[24]	LIU H R, WU G, ZHOU B, et al. Structure and function of cholesteryl ester transfer protein in the tree shrew[J]. Lipids, 2011, 46(7):607-616. DOI: 10.1007/s11745-011-3552-2 .
[25]	ZHENG L T, CHEN S Y, WU Q L, et al. Tree shrews as a new animal model for systemic sclerosis research[J]. Front Immunol, 2024, 15:1315198. DOI: 10.3389/fimmu.2024.1315198 .
[26]	KOTHARI M, WANJARI A, ACHARYA S, et al. A comprehensive review of monoclonal antibodies in modern medicine: tracing the evolution of a revolutionary therapeutic approach[J]. Cureus, 2024, 16(6): e61983. DOI: 10.7759/cureus.61983 .
[27]	LITTLE M, KIPRIYANOV S M, LE GALL F, et al. Of mice and men: hybridoma and recombinant antibodies[J]. Immunol Today, 2000, 21(8):364-370. DOI: 10.1016/s0167-5699(00)01668-6 .
[28]	SKOWICKI M, LIPIŃSKI T. The development of methods for obtaining monoclonal antibody-producing cells[J]. Postepy Hig Med Dosw, 2016, 70:367-379. DOI: 10.5604/1732269 3.1200552 .
[29]	邝贞结. 基因工程重组抗体技术的研究进展[J]. 广东畜牧兽医科技, 2010, 35(5):3-6. DOI: 10.3969/j.issn.1005-8567.2010.05.001 .
	KUANG Z J. Research progress of recombinant antibody technology in genetic engineering[J]. Guangdong J Anim Vet Sci, 2010, 35(5):3-6. DOI: 10.3969/j.issn.1005-8567.2010.05.001 .
[30]	董新莹, 高晓薇, 宋浩, 等. 纳米抗体的研究进展及其应用现状[J]. 生物工程学报, 2024, 40(12):4324-4338. DOI: 10.13345/j.cjb.240366 .
	DONG X Y, GAO X W, SONG H, et al. Research progress and application of nanobodies[J]. Chin J Biotechnol, 2024, 40(12):4324-4338. DOI: 10.13345/j.cjb.240366 .
[31]	武瑞君, 桑晓冬, 李治非, 等. 抗体技术的研发现状与展望[J]. 中国药理学与毒理学杂志, 2021, 35(5):374-381. DOI: 10.3867/j.issn.1000-3002.2021.05.007 .
	WU R J, SANG X D, LI Z F, et al. Development and prospect of antibody technology[J]. Chin J Pharmacol Toxicol, 2021, 35(5):374-381. DOI: 10.3867/j.issn.1000-3002.2021.05.007 .
[32]	GILEADI O. Recombinant protein expression in E. coli: a historical perspective[J]. Methods Mol Biol, 2017, 1586:3-10. DOI: 10.1007/978-1-4939-6887-9_1 .
[33]	ROSANO G L, CECCARELLI E A. Recombinant protein expression in Escherichia coli: advances and challenges[J]. Front Microbiol, 2014, 5:172. DOI: 10.3389/fmicb.2014.00172 .
[34]	SUN J M, GUO X P, YU P, et al. Vasorin deficiency leads to cardiac hypertrophy by targeting MYL7 in young mice[J]. J Cell Mol Med, 2022, 26(1):88-98. DOI: 10.1111/jcmm.17034 .
[35]	SUN J M, YIN S W, LI Q R, et al. VASN knockout induces myocardial fibrosis in mice by downregulating non-collagen fibers and promoting inflammation[J]. Front Pharmacol, 2025, 15:1500617. DOI: 10.3389/fphar.2024.1500617 .

树鼩Vasorin单克隆抗体的制备及其应用探索

Preparation of Monoclonal Antibody to Vasorin in Tree Shrew and Exploration of Its Application

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