肺癌蛋白芯片检测系统在乳腺癌诊断中的价值
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摘要
目前,癌症依然是当今世界的难题,随着生态环境的恶化和生活节奏的加快,癌症在人群中的发病率逐年增高,而早期诊断和早期治疗已经被医学界认为是解决癌症最有效的方法。肿瘤标志物是反映肿瘤存在和生长的一类物质,通过肿瘤标志物相关参数的变化可以获得大量的肿瘤生长信息,也为早期癌症诊断提供了依据。在临床诊断中有通用的肿瘤标志物,这种标志物往往在多个癌症表达异常,我们可以结合几个通用标志物通过检测表达量的变化来判断癌症的种类。钟理博士所构建的新型肺癌肿瘤标志物蛋白芯片检测系统对于检测非小细胞肺癌有着很好的效果,但是该检测系统对于其它癌症的诊断价值还未见报道。新型蛋白芯片筛选出来的肺癌肿瘤标志物里面很有可能含有能检测其它癌症的肿瘤标志物,一旦发现可以大大提升该系统的应用价值。
对肺癌T7噬菌体展示文库进行生物淘洗,从而富集了大量肺癌免疫原性噬菌体,再通过蛋白芯片的高通量筛选获得了94个肺癌肿瘤标志物。将这些标志物制备在醛基化处理玻片上构建成为诊断芯片,并用15份乳腺癌血清和15份正常人血清处理芯片,对该诊断芯片进行统计学分析和诊断价值的评估。t检验结果显示只有编号为573的噬菌体克隆的P值较小,为0.0526。虽然标志物差异水平没有达到显著标准,但是ROC曲线分析结果显示,573号噬菌体克隆拥有不错的诊断价值,ROC曲线下面积达到0.716，敏感性和特异性最高为70%,很有可能是特异性的肿瘤标志物。运用多肿瘤标志物联合检测ROC曲线下面积虽有提高但是不明显。下一步需要对573号克隆做进一步的研究和鉴定。
构建一个成熟的蛋白芯片诊断体系需要大量的时间和经费,利用已有的肺癌诊断系统对于其它癌症进行检测是一个快速、经济的方法,与此同时,我们也了解两种癌症的肿瘤标志物之间的区别,有助于我们开发多肿瘤标志物检测系统。
Cancer is one of the biggest problems in the world. Its incidence becomes higher with the deterioration of environment and the change of life style. The early diagnose and therapy is the best way against cancer. Tumor markers could reflect the presence and growth of tumor, by which, a lot of information of tumor can be obtained and the early diagnose gist can be provided. Now, there are some current tumor marker in the clinical diagnosis. We can judge the style of cancer through the changes of expression combining with several current markers. The new multiple tumor marker protein chip detective system which was adopted by Doctor Zhong is valuable for detection of non small cell lung cancer, but the diagnostic value for the other cancers have not reported. It maybe has tumor marker which could detect the other cancers in lung cancer tumor marker that are screened from new protein chip system. It will highly improve the application value of this system when be proved.
After several biopanning which screened from lung cancer T7 phage display library, we enrichment lots of immunogenicity bacteriophage of lung cancer, and we can got 94 lung cancer tumor markers by high-throughput screening of protein chips. These markers was put on aldehyde glass slide to construct detection chips. These chips hybridized with 15 breast cancer serums and 15 normal serums we collected. The data from expriment analyzed statistically and assessed its diagnostic value. The result showed that only No.573 phage clone's P value is smaller, that is 0.0526. Although,the different level of No.573 can not reach significant differences, the ROC curves results indicates that No.537 bacteriophage has better diagnostic value, the area under ROC curves is nearly 0.716, sensitivity and specificity reach 70%, it maybe the tumor marker with specificity. Although it is improved for combined detection of ROC curve by different tumor markers, the result is still not obvious. Next we want to reseach deeply and identify No.573 phage clone.
It needs a lot of time and funds to construct a mature protein chip diagnosis system, so making use of existing lung cancer diagnosis system to make advisable diagnostic analysis is more faster and economical. Meanwhile, we can get some information about difference tumor marker about the two kinds of cancer, it will give some help to develop multi-tumor marker detective system.
对肺癌T7噬菌体展示文库进行生物淘洗,从而富集了大量肺癌免疫原性噬菌体,再通过蛋白芯片的高通量筛选获得了94个肺癌肿瘤标志物。将这些标志物制备在醛基化处理玻片上构建成为诊断芯片,并用15份乳腺癌血清和15份正常人血清处理芯片,对该诊断芯片进行统计学分析和诊断价值的评估。t检验结果显示只有编号为573的噬菌体克隆的P值较小,为0.0526。虽然标志物差异水平没有达到显著标准,但是ROC曲线分析结果显示,573号噬菌体克隆拥有不错的诊断价值,ROC曲线下面积达到0.716，敏感性和特异性最高为70%,很有可能是特异性的肿瘤标志物。运用多肿瘤标志物联合检测ROC曲线下面积虽有提高但是不明显。下一步需要对573号克隆做进一步的研究和鉴定。
构建一个成熟的蛋白芯片诊断体系需要大量的时间和经费,利用已有的肺癌诊断系统对于其它癌症进行检测是一个快速、经济的方法,与此同时,我们也了解两种癌症的肿瘤标志物之间的区别,有助于我们开发多肿瘤标志物检测系统。
Cancer is one of the biggest problems in the world. Its incidence becomes higher with the deterioration of environment and the change of life style. The early diagnose and therapy is the best way against cancer. Tumor markers could reflect the presence and growth of tumor, by which, a lot of information of tumor can be obtained and the early diagnose gist can be provided. Now, there are some current tumor marker in the clinical diagnosis. We can judge the style of cancer through the changes of expression combining with several current markers. The new multiple tumor marker protein chip detective system which was adopted by Doctor Zhong is valuable for detection of non small cell lung cancer, but the diagnostic value for the other cancers have not reported. It maybe has tumor marker which could detect the other cancers in lung cancer tumor marker that are screened from new protein chip system. It will highly improve the application value of this system when be proved.
After several biopanning which screened from lung cancer T7 phage display library, we enrichment lots of immunogenicity bacteriophage of lung cancer, and we can got 94 lung cancer tumor markers by high-throughput screening of protein chips. These markers was put on aldehyde glass slide to construct detection chips. These chips hybridized with 15 breast cancer serums and 15 normal serums we collected. The data from expriment analyzed statistically and assessed its diagnostic value. The result showed that only No.573 phage clone's P value is smaller, that is 0.0526. Although,the different level of No.573 can not reach significant differences, the ROC curves results indicates that No.537 bacteriophage has better diagnostic value, the area under ROC curves is nearly 0.716, sensitivity and specificity reach 70%, it maybe the tumor marker with specificity. Although it is improved for combined detection of ROC curve by different tumor markers, the result is still not obvious. Next we want to reseach deeply and identify No.573 phage clone.
It needs a lot of time and funds to construct a mature protein chip diagnosis system, so making use of existing lung cancer diagnosis system to make advisable diagnostic analysis is more faster and economical. Meanwhile, we can get some information about difference tumor marker about the two kinds of cancer, it will give some help to develop multi-tumor marker detective system.
引文
[1]朱赓伯.癌症发病机理新说.家庭医学,2006(1)：35.
[2]Douglas Hanahan, Robert A. Weinberg. The Hallmarks of Cancer. Cell,2000,100:57-70.
[3]柳青.全球影响癌症发病的9种行为与环境危险因素.中华医学杂志,2006,86(1)：44.
[4]成书均.肿瘤早期诊断技术及标准研讨会纪要.中国医药导刊,2006(4)：86.
[5]孙燕.肿瘤的发生情况与防治趋向.中国医学论坛报,2002(14).
[6]焦顺昌.生活方式与肿瘤预防.解放军保健医学杂志,2005,7(2)：72-74.
[7]王晓,曾纪权,欧阳江.癌症控制防大于治.医学与哲学,2006,27(1)：13-14.
[8]Samir Hanash. Integrated global profiling of cancer. Nature,2004,4:638-643.
[9]Hanash SM, Bobek MP, Rickman DS, et al. Puravs E. Integrating cancer genomics and proteomics in the post-genome era. Proteomics,2002,2:69-75.
[10]Srinivas PR, Kramer BS, Srivastava S. Trends in biomarker research for cancer detection. Lancet Oncol,2001,2:698-704.
[11]袁玫.实体肿瘤标志研究进展.实用肿瘤杂志,1994,9(4)：251-253.
[12]李春海.肿瘤标志的基础与临床应用.中华医学信息导报,2004,19(17)：20.
[13]马乃绪.肿瘤标志物的检测及临床应用.前卫医药杂志,2001,18(4)：314-317.
[14]陈燕.肿瘤标志物临床应用原则.医学检验与临床,2006,17(1)：1-2.
[15]李嘉,刘善超,刘英菊,等.肿瘤标志物与诊断技术.大学化学,2004,19(1)：1-9.
[16]贺宇.肿瘤标志物临床应用研究现状.实用医技杂志,2006,13(7)：1205-1206.
[17]管梁,陈刚,朱承谟.肿瘤标志物的实验诊断及其进展.标记免疫分析与临床,2001,8(4)：228-233.
[18]Greg L. Perkins, Evan D. Slater, Georganne K. Sanders, et al. Serum Tumor Markers. American Family Physician,2003,6:1075-1085.
[19]Fleisher M, Dnistrian AM, Sturgeon CM, et al. Practice guidelines and recommendations for use tumor markers in the clinic. Washington DC:AACC Press,2002:20-25.
[20]M.B. Gretzer, A.W. Partin. PSA markers in prostate cancer detection. Urol. Clin. North Am,2003, 30:677.
[21]季晨阳.乳腺癌肿瘤标志物的临床应用和研究进展.实用医学杂志,2000,16(7)：521-523.
[22]刘春,范卫华.肺癌肿瘤标志物.山东医药,2006,46(11)：65.
[23]张品良,邵松.乳腺癌肿瘤标志物.山东医药,2006,46(11)：65-67.
[24]武建国.肿瘤标志物的若干进展.临床检验杂志,2002,20(特刊)：33-38.
[25]顾克东,张雅青.肿瘤标志物的最新研究技术及其临床检测方法.西北民族大学学报,2005,26(59)：86-88.
[26]Nilofer S. Azad, Nabila Rasool, Christina M. Annunziata, et al. Proteomics in Clinical Trials and Practice Molecular & Cellular Proteomics,2006,5:1819-1829.
[27]Hoehn G. T., Suffredini, A. F. Proteomics. Crit. Care Med,2005,33(suppl.):444-448.
[28]Choi KS, Song L, Park YM, et al. Analysis of human plasma proteome by 2DE- and 2D nanoLC-based mass spectrometry. Prep Biochem Biotechnol,2006,36:3217.
[29]Klade CS, Voss T, Krystek E, et al. Identification of tumor antigens in renal cell carcinoma by serological proteome analysis. Proteomics,2001,1:888-890.
[30]Naour FL, Brichory F, Beretta L, et al. Identification of tumor-associated antigens using proteomics. Technol Cancer Res Treat,2002,1:257-262.
[31]Le Naour F, Brichory F, Misek DE, et al. A distinct repertoire of autoantibodies in hepatocellular carcinoma identified by proteomic analysis. Mol Cell Proteomics,2002,1:197-203.
[32]Feng JT, Liu YK, Song HY, et al. Heat shock protein 27:a potential biomarker for hepatocellular carcinoma identified by serum proteome analysis. Proteomics,2005,5:4581-4588.
[33]Ji Gaoa, Leah-Ann Garulacanb, Stephen M. Storma. Biomarker discovery in biological fluids. Methods,2005,35:291-302.
[34]容毓,邓粤敏.几种肿瘤标志物检测方法的进展及应用.实用医学杂志,2006,22(9)：1094-1095.
[35]张青云.肿瘤标志物临床检测技术现状及发展趋势.中华检验医学杂志,2006,29(7)：585-589.
[36]杨涛,杨金亮,魏于全.SELDI蛋白芯片技术及其应用研究进展.肿瘤防治杂志,2004,11(10)：1101-1103.
[37]Wright G L, Cazares L H, Leung S M, et al. SELDI mass spectrometry:a novel protein biochip technology for dectection of biomarker in complex protein mixtures. Prostate Cancer and Prostatic Disease,1999,2 (5):264-276.
[38]Wlad Kusnezow, J&rg D, Hoheisel, et al. Antibody Microarrays:Promises and Problems. Biotechniques,2002,10 (33):14-23.
[39]Yoon S. Cho-Chung. Autoantibody biomarkers in the detection of cancer. Biochimica et Biophysica Acta 1762,2006:587-591.
[40]Hailing Lu, Vivian Goodell, Mary L Disis. Targeting serum antibody for cancer diagnosis:a focus on colorectal cancer. Expert Opin Ther Targets,2007, 11(2):235-244.
[41]Li Zhong, Giovanna E. Hidalgo, Arnold J. Stromberg. Using protein microarray as a diagnostic assay for non-small cell lung cancer. American journal of respiratory and critical care medicine,2005, 172:1-7.
[42]Smit h G P. Filamentous fusion phage:novel expression vectors that display cloned antigens on the virion surface. Science,1985,228:1315-1317.
[43]Pandey A, Mann M. Proteomics to study genes and genomes. Nature,2002,405(15):837-846.
[44]Cheng J, Zhong Y W, Liu Y, et al. Screening and identification of a humanized single chain variable region antibody for hepatitis C virus non-structural 3 protein. Chin J Immunol,2000,16 (4):246.
[45]袁改玲,郭媛华,张俊英,等.噬菌体表面展示技术的发展及应用.动物医学进展,2005,26(3)：1-4.
[46]张忠东,成军,张树林.噬菌体展示技术的原理及应用.世界华人消化杂志，2003,11(4)：459-461.
[47]Doorbar J, Winter G. Isolation of a peptide antagonist to thethrombin receptor using phage display. J Mol Biol,1994,244:361-369.
[48]Pasqualini R, Ruoslahti E. Organ targeting in vivo using phage display peptide libraries. Nature,1996, 380:364-366.
[49]Clackson T, Hoogenboom HR, Griffiths AD, et al. Making antibody fragments using phage display libraries. Nature,1991,352:624-628.
[50]Tang Y, Jiang N, Parakh C, et al. Selection of linkers for acatalytic single chain antibody using phage display technology. J Biol Chem,1996,271:15682-15686.
[51]Rhyner C, Kodzius R, Crameri R. Direct selection of cDNAs from filamentous phage surface display libraries:potential and limitations. Curr Pharm,2002,3:13-21.
[52]李丽芳,张映.噬菌体展示文库的筛选技术.生物技术通报,2005(4)：36-38.
[53]Gygi S P, Rochon Y, Franza B, Correlation between protein and mRNA abundance in yeast, et al. Mol Cell Biol,1999,19(3):1720.
[54]Walter G, Bussow K, Cahill D, et al. Protein arrays for gene expression and molecular interaction screening. Curr Opin Microbiol,2000,3(3):298-302.
[55]MacBeath G, Schreiber SL. Printing proteins as microarrays for high-throughput function determination. Science,2000,289 (5485):17602-17631.
[56]石志红,张新祥,常文保.蛋白芯片研究进展.大学化学,2002,17(3)：1-5.
[57]陈玮莹,温博贵.蛋白芯片的基本原理及技术研究现状.国外医学临床生物化学与检验学分册,2002,23(1)：5-6.
[58]Zhu H, Klemic J F, Chang S, et al. Analysis of yeast protein kinases using protein chips. Nat Genet, 2000,26(3):283-289.
[59]Cahill D J. Protein and antibody arrays and their medical applications. Journal of Immunological Methods,2001,250:81-91.
[60]高志贤.蛋白芯片技术研究进展及其应用.现代仪器,2004(3)：1-5.
[61]Kumble K D. Protein microarrays:new tools for pharmaceutical development. Anal Bioanal Chem, 2003,377:812-819.
[62]余章斌.蛋白质芯片检测技术的研究进展.国外医学临床生物化学与检验学分册,2005,26(11)：841-843.
[63]Yu W, Gu N, Zhang HQ, et al. Microarray preparation based on oxidation of agarose gel and subsequent enzyme immunoassay. Sensors and Actuators B:Chemical,2004,98(1):83-91.
[64]靳刚,王战会.光学蛋白质芯片.自然杂志,2001,23(5)：286-289.
[65]Shiwa M, Nishimura Y, Wakatabe R, et al. Rapid discovery and identification of a tissue specific tumor biomarker from 39 human cancer cell lines using the SELDI Protein Chip platform. Biochem Biophys Res Commun,2003,309(1):18-25.
[66]Kaur J, Singh KV, Schmid A H, et al. Atomic force spectroscopy-based study of antibody pesticide interactions for characterization of immunosensor surface. Biosens Bioelectron,2004,20(2):284-293.
[67]Lee JW, Sim SJ, Cho SM, et al. Characterization of a self-assembled monolayer of thiol on a gold surface and the fabrication of a biosensor chip based on surface plasmon resonance for detecting anti-GAD antibody. Biosens Bioelectron,2005,20 (7):1422-1427.
[2]Douglas Hanahan, Robert A. Weinberg. The Hallmarks of Cancer. Cell,2000,100:57-70.
[3]柳青.全球影响癌症发病的9种行为与环境危险因素.中华医学杂志,2006,86(1)：44.
[4]成书均.肿瘤早期诊断技术及标准研讨会纪要.中国医药导刊,2006(4)：86.
[5]孙燕.肿瘤的发生情况与防治趋向.中国医学论坛报,2002(14).
[6]焦顺昌.生活方式与肿瘤预防.解放军保健医学杂志,2005,7(2)：72-74.
[7]王晓,曾纪权,欧阳江.癌症控制防大于治.医学与哲学,2006,27(1)：13-14.
[8]Samir Hanash. Integrated global profiling of cancer. Nature,2004,4:638-643.
[9]Hanash SM, Bobek MP, Rickman DS, et al. Puravs E. Integrating cancer genomics and proteomics in the post-genome era. Proteomics,2002,2:69-75.
[10]Srinivas PR, Kramer BS, Srivastava S. Trends in biomarker research for cancer detection. Lancet Oncol,2001,2:698-704.
[11]袁玫.实体肿瘤标志研究进展.实用肿瘤杂志,1994,9(4)：251-253.
[12]李春海.肿瘤标志的基础与临床应用.中华医学信息导报,2004,19(17)：20.
[13]马乃绪.肿瘤标志物的检测及临床应用.前卫医药杂志,2001,18(4)：314-317.
[14]陈燕.肿瘤标志物临床应用原则.医学检验与临床,2006,17(1)：1-2.
[15]李嘉,刘善超,刘英菊,等.肿瘤标志物与诊断技术.大学化学,2004,19(1)：1-9.
[16]贺宇.肿瘤标志物临床应用研究现状.实用医技杂志,2006,13(7)：1205-1206.
[17]管梁,陈刚,朱承谟.肿瘤标志物的实验诊断及其进展.标记免疫分析与临床,2001,8(4)：228-233.
[18]Greg L. Perkins, Evan D. Slater, Georganne K. Sanders, et al. Serum Tumor Markers. American Family Physician,2003,6:1075-1085.
[19]Fleisher M, Dnistrian AM, Sturgeon CM, et al. Practice guidelines and recommendations for use tumor markers in the clinic. Washington DC:AACC Press,2002:20-25.
[20]M.B. Gretzer, A.W. Partin. PSA markers in prostate cancer detection. Urol. Clin. North Am,2003, 30:677.
[21]季晨阳.乳腺癌肿瘤标志物的临床应用和研究进展.实用医学杂志,2000,16(7)：521-523.
[22]刘春,范卫华.肺癌肿瘤标志物.山东医药,2006,46(11)：65.
[23]张品良,邵松.乳腺癌肿瘤标志物.山东医药,2006,46(11)：65-67.
[24]武建国.肿瘤标志物的若干进展.临床检验杂志,2002,20(特刊)：33-38.
[25]顾克东,张雅青.肿瘤标志物的最新研究技术及其临床检测方法.西北民族大学学报,2005,26(59)：86-88.
[26]Nilofer S. Azad, Nabila Rasool, Christina M. Annunziata, et al. Proteomics in Clinical Trials and Practice Molecular & Cellular Proteomics,2006,5:1819-1829.
[27]Hoehn G. T., Suffredini, A. F. Proteomics. Crit. Care Med,2005,33(suppl.):444-448.
[28]Choi KS, Song L, Park YM, et al. Analysis of human plasma proteome by 2DE- and 2D nanoLC-based mass spectrometry. Prep Biochem Biotechnol,2006,36:3217.
[29]Klade CS, Voss T, Krystek E, et al. Identification of tumor antigens in renal cell carcinoma by serological proteome analysis. Proteomics,2001,1:888-890.
[30]Naour FL, Brichory F, Beretta L, et al. Identification of tumor-associated antigens using proteomics. Technol Cancer Res Treat,2002,1:257-262.
[31]Le Naour F, Brichory F, Misek DE, et al. A distinct repertoire of autoantibodies in hepatocellular carcinoma identified by proteomic analysis. Mol Cell Proteomics,2002,1:197-203.
[32]Feng JT, Liu YK, Song HY, et al. Heat shock protein 27:a potential biomarker for hepatocellular carcinoma identified by serum proteome analysis. Proteomics,2005,5:4581-4588.
[33]Ji Gaoa, Leah-Ann Garulacanb, Stephen M. Storma. Biomarker discovery in biological fluids. Methods,2005,35:291-302.
[34]容毓,邓粤敏.几种肿瘤标志物检测方法的进展及应用.实用医学杂志,2006,22(9)：1094-1095.
[35]张青云.肿瘤标志物临床检测技术现状及发展趋势.中华检验医学杂志,2006,29(7)：585-589.
[36]杨涛,杨金亮,魏于全.SELDI蛋白芯片技术及其应用研究进展.肿瘤防治杂志,2004,11(10)：1101-1103.
[37]Wright G L, Cazares L H, Leung S M, et al. SELDI mass spectrometry:a novel protein biochip technology for dectection of biomarker in complex protein mixtures. Prostate Cancer and Prostatic Disease,1999,2 (5):264-276.
[38]Wlad Kusnezow, J&rg D, Hoheisel, et al. Antibody Microarrays:Promises and Problems. Biotechniques,2002,10 (33):14-23.
[39]Yoon S. Cho-Chung. Autoantibody biomarkers in the detection of cancer. Biochimica et Biophysica Acta 1762,2006:587-591.
[40]Hailing Lu, Vivian Goodell, Mary L Disis. Targeting serum antibody for cancer diagnosis:a focus on colorectal cancer. Expert Opin Ther Targets,2007, 11(2):235-244.
[41]Li Zhong, Giovanna E. Hidalgo, Arnold J. Stromberg. Using protein microarray as a diagnostic assay for non-small cell lung cancer. American journal of respiratory and critical care medicine,2005, 172:1-7.
[42]Smit h G P. Filamentous fusion phage:novel expression vectors that display cloned antigens on the virion surface. Science,1985,228:1315-1317.
[43]Pandey A, Mann M. Proteomics to study genes and genomes. Nature,2002,405(15):837-846.
[44]Cheng J, Zhong Y W, Liu Y, et al. Screening and identification of a humanized single chain variable region antibody for hepatitis C virus non-structural 3 protein. Chin J Immunol,2000,16 (4):246.
[45]袁改玲,郭媛华,张俊英,等.噬菌体表面展示技术的发展及应用.动物医学进展,2005,26(3)：1-4.
[46]张忠东,成军,张树林.噬菌体展示技术的原理及应用.世界华人消化杂志，2003,11(4)：459-461.
[47]Doorbar J, Winter G. Isolation of a peptide antagonist to thethrombin receptor using phage display. J Mol Biol,1994,244:361-369.
[48]Pasqualini R, Ruoslahti E. Organ targeting in vivo using phage display peptide libraries. Nature,1996, 380:364-366.
[49]Clackson T, Hoogenboom HR, Griffiths AD, et al. Making antibody fragments using phage display libraries. Nature,1991,352:624-628.
[50]Tang Y, Jiang N, Parakh C, et al. Selection of linkers for acatalytic single chain antibody using phage display technology. J Biol Chem,1996,271:15682-15686.
[51]Rhyner C, Kodzius R, Crameri R. Direct selection of cDNAs from filamentous phage surface display libraries:potential and limitations. Curr Pharm,2002,3:13-21.
[52]李丽芳,张映.噬菌体展示文库的筛选技术.生物技术通报,2005(4)：36-38.
[53]Gygi S P, Rochon Y, Franza B, Correlation between protein and mRNA abundance in yeast, et al. Mol Cell Biol,1999,19(3):1720.
[54]Walter G, Bussow K, Cahill D, et al. Protein arrays for gene expression and molecular interaction screening. Curr Opin Microbiol,2000,3(3):298-302.
[55]MacBeath G, Schreiber SL. Printing proteins as microarrays for high-throughput function determination. Science,2000,289 (5485):17602-17631.
[56]石志红,张新祥,常文保.蛋白芯片研究进展.大学化学,2002,17(3)：1-5.
[57]陈玮莹,温博贵.蛋白芯片的基本原理及技术研究现状.国外医学临床生物化学与检验学分册,2002,23(1)：5-6.
[58]Zhu H, Klemic J F, Chang S, et al. Analysis of yeast protein kinases using protein chips. Nat Genet, 2000,26(3):283-289.
[59]Cahill D J. Protein and antibody arrays and their medical applications. Journal of Immunological Methods,2001,250:81-91.
[60]高志贤.蛋白芯片技术研究进展及其应用.现代仪器,2004(3)：1-5.
[61]Kumble K D. Protein microarrays:new tools for pharmaceutical development. Anal Bioanal Chem, 2003,377:812-819.
[62]余章斌.蛋白质芯片检测技术的研究进展.国外医学临床生物化学与检验学分册,2005,26(11)：841-843.
[63]Yu W, Gu N, Zhang HQ, et al. Microarray preparation based on oxidation of agarose gel and subsequent enzyme immunoassay. Sensors and Actuators B:Chemical,2004,98(1):83-91.
[64]靳刚,王战会.光学蛋白质芯片.自然杂志,2001,23(5)：286-289.
[65]Shiwa M, Nishimura Y, Wakatabe R, et al. Rapid discovery and identification of a tissue specific tumor biomarker from 39 human cancer cell lines using the SELDI Protein Chip platform. Biochem Biophys Res Commun,2003,309(1):18-25.
[66]Kaur J, Singh KV, Schmid A H, et al. Atomic force spectroscopy-based study of antibody pesticide interactions for characterization of immunosensor surface. Biosens Bioelectron,2004,20(2):284-293.
[67]Lee JW, Sim SJ, Cho SM, et al. Characterization of a self-assembled monolayer of thiol on a gold surface and the fabrication of a biosensor chip based on surface plasmon resonance for detecting anti-GAD antibody. Biosens Bioelectron,2005,20 (7):1422-1427.