Biology:Antibody microarray

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Samples of antibody microarray creations and detections.

An antibody microarray (also known as antibody array) is a specific form of protein microarray. In this technology, a collection of captured antibodies are spotted and fixed on a solid surface such as glass, plastic, membrane, or silicon chip, and the interaction between the antibody and its target antigen is detected. Antibody microarrays are often used for detecting protein expression from various biofluids including serum, plasma and cell or tissue lysates. Antibody arrays may be used for both basic research and medical and diagnostic applications.[1][2][3][4]

Background

The concept and methodology of antibody microarrays were first introduced by Tse Wen Chang in 1983 in a scientific publication[5] and a series of patents,[6][7][8] when he was working at Centocor in Malvern, Pennsylvania. Chang coined the term “antibody matrix” and discussed “array” arrangement of minute antibody spots on small glass or plastic surfaces. He demonstrated that a 10×10 (100 in total) and 20×20 (400 in total) grid of antibody spots could be placed on a 1×1 cm surface. He also estimated that if an antibody is coated at a 10 μg/mL concentration, which is optimal for most antibodies, 1 mg of antibody can make 2,000,000 dots of 0.25 mm diameter. Chang's invention focused on the employment of antibody microarrays for the detection and quantification of cells bearing certain surface antigens, such as CD antigens and HLA allotypic antigens, particulate antigens, such as viruses and bacteria, and soluble antigens. The principle of "one sample application, multiple determinations", assay configuration, and mechanics for placing absorbent dots described in the paper and patents should be generally applicable to different kinds of microarrays. When Tse Wen Chang and Nancy T. Chang were setting up Tanox, Inc. in Houston, Texas in 1986, they purchased the rights on the antibody matrix patents from Centocor as part of the technology base to build their new startup. Their first product in development was an assay, termed “immunosorbent cytometry”,[9] which could be employed to monitor the immune status, i.e., the concentrations and ratios of CD3+, CD4+, and CD8+ T cells, in the blood of HIV-infected individuals.

The theoretical background for protein microarray-based ligand binding assays was further developed by Roger Ekins and colleagues in the late 1980s.[10][11][12] According to the model, antibody microarrays would not only permit simultaneous screening of an analyte panel, but would also be more sensitive and rapid than conventional screening methods. Interest in screening large protein sets only arose as a result of the achievements in genomics by DNA microarrays and the Human Genome Project.

The first array approaches attempted to miniaturize biochemical and immunobiological assays usually performed in 96-well microtiter plates. While 96-well plate-based antibody arrays have high-throughput capability, the small surface area in each well limits the number of antibody spots and thus, the number of analytes detected. Other solid supports, such as glass slides and nitrocellulose membranes, were subsequently utilized to develop arrays which could accommodate larger panels of antibodies.[13] Nitrocellulose membrane-based arrays are flexible, easy to handle, and have increased protein binding capacity, but are less amenable to high throughput or automated processing. Chemically derivatized glass slides allow for printing of sub-microliter sized antibody spots, reducing the array surface area without sacrificing spot density. This in turn reduces the volume of sample consumed. Glass slide-based arrays, owing to their smooth and rigid structure, can also be easily fitted to high-throughput liquid handling systems.

Most antibody array systems employ 1 of 2 non-competitive methods of immunodetection: single-antibody (label-based) detection and 2-antibody (sandwich-based) detection. The latter method, in which analyte detection requires the binding of 2 distinct antibodies (a capture antibody and a reporter antibody, each binding to a unique epitope), confers greater specificity and lower background signal compared with label-based immunodetection (where only 1 capture antibody is used and detection is achieved by chemically labeling all proteins in the starting sample). Sandwich-based antibody arrays usually attain the highest specificity and sensitivity (ng – pg levels) of any array format; their reproducibility also enables quantitative analysis to be performed.[14][15] Due to the difficulty of developing matched antibody pairs that are compatible with all other antibodies in the panel, small arrays often make use of a sandwich approach. Conversely, high-density arrays are easier to develop at a lower cost using the single antibody label-based approach. In this methodology, one set of specific antibodies is used and all the proteins in a sample are labelled directly by fluorescent dyes or haptens.

Initial uses of antibody-based array systems included detecting IgGs and specific subclasses,[16][17] analyzing antigens,[18] screening recombinant antibodies,[19][20] studying yeast protein kinases,[21] analyzing autoimmune antibodies,[22] and examining protein-protein interactions.[23][24][25] The first approach to simultaneously detect multiple cytokines from physiological samples using antibody array technology was by Ruo-Pan Huang and colleagues in 2001.[26] Their approach used Hybond ECL membranes to detect a small panel of 24 cytokines from cell culture conditioned media and patient's sera and was able to profile cytokine expression at physiological levels. Huang took this technology and started a new business, RayBiotech, Inc., the first to successfully commercialize a planar antibody array.

In the last ten years, the sensitivity of the method was improved by an optimization of the surface chemistry as well as dedicated protocols for their chemical labeling.[27] Currently, the sensitivity of antibody arrays is comparable to that of ELISA[28][29] and antibody arrays are regularly used for profiling experiments on tissue samples, plasma or serum samples and many other sample types. One main focus in antibody array based profiling studies is biomarker discovery, specifically for cancer.[30][31][32][33][34] For cancer-related research, the development and application of an antibody array comprising 810 different cancer-related antibodies was reported in 2010.[35] Also in 2010, an antibody array comprising 507 cytokines, chemokines, adipokines, growth factors, angiogenic factors, proteases, soluble receptors, soluble adhesion molecules, and other proteins was used to screen the serum of ovarian cancer patients and healthy individuals and found a significant difference in protein expression between normal and cancer samples.[36] More recently, antibody arrays have helped determine specific allergy-related serum proteins whose levels are associated with glioma and can reduce the risk years before diagnosis.[37] Protein profiling with antibody arrays have also proven successful in areas other than cancer research, specifically in neurological diseases such as Alzheimer's. A number of studies have attempted to identify biomarker panels that can distinguish Alzheimer's patients, and many have used antibody arrays in this process. Jaeger and colleagues measured nearly 600 circulatory proteins to discover biological pathways and networks affected in Alzheimer's and explored the positive and negative relationships of the levels of those individual proteins and networks with the cognitive performance of Alzheimer's patients.[38] Currently the largest commercially available sandwich-based antibody array detects 1000 different proteins.[39] In addition, antibody microarray based protein profiling services are available analyzing protein abundance and protein phosphorylation or ubiquitinylation status of 1030 proteins in parallel.[40]

Antibody arrays are often used for detecting protein expression from many sample types, but also in those with various preparations. Jiang and colleagues illustrated nicely the correlation between array protein expression in two different blood preparations: serum and dried blood spots.[41] These different blood sample preparations were analyzed using three antibody array platforms: sandwich-based, quantitative, and label-based, and a strong correlation in protein expression was found, suggesting that dried blood spots, which are a more convenient, safe, and inexpensive means of obtaining blood especially in non-hospitalized public health areas, can be used effectively with antibody array analysis for biomarker discovery, protein profiling, and disease screening, diagnosis, and treatment.

Applications

Using antibody microarray in different medical diagnostic areas has attracted researchers attention. Digital bioassay is an example of such research domains. In this technology, an array of microwells on a glass/polymer chip are seeded with magnetic beads (coated with fluorescent tagged antibodies), subjected to targeted antigens and then characterised by a microscope through counting fluorescing wells. A cost-effective fabrication platform (using OSTE polymers) for such microwell arrays has been recently demonstrated and the bio-assay model system has been successfully characterised.[42] Furthermore, immunoassays on thiol-ene "synthetic paper" micropillar scaffolds have shown to generate a superior fluorescence signal.[43]

See also

References

  1. ↑ "A 200-antibody microarray biochip for environmental monitoring: searching for universal microbial biomarkers through immunoprofiling". Anal. Chem. 80 (21): 7970–9. November 2008. doi:10.1021/ac8008093. PMID 18837515. 
  2. ↑ Chaga GS (2008). "Antibody Arrays for Determination of Relative Protein Abundances". Tissue Proteomics. Methods in Molecular Biology. 441. pp. 129–51. doi:10.1007/978-1-60327-047-2_9. ISBN 978-1-58829-679-5. 
  3. ↑ Wilson J. J.Expression error: Unrecognized word "et". (2015). Chapter Seven-Antibody Arrays in Biomarker Discovery. 69. 255–324. doi:10.1016/bs.acc.2015.01.002. ISBN 9780128022658. 
  4. ↑ Lin Y., Huang R.C., Cao X., Wang S.-M., Shi Q., Huang R.-P. (2003). "Detection of multiple cytokines by protein arrays from cell lysate and tissue lysate". Clin Chem Lab Med 41 (2): 139–145. doi:10.1515/cclm.2003.023. PMID 12666998. 
  5. ↑ Chang TW (December 1983). "Binding of cells to matrixes of distinct antibodies coated on solid surface". J. Immunol. Methods 65 (1–2): 217–23. doi:10.1016/0022-1759(83)90318-6. PMID 6606681. 
  6. ↑ Chang, Tse W. U.S. Patent 4,591,570 "Matrix of antibody-coated spots for determination of antigens", Priority date February 2, 1983
  7. ↑ Chang, Tse W. U.S. Patent 4,829,010 "Immunoassay device enclosing matrixes of antibody spots for cell determinations", Priority date March 13, 1987
  8. ↑ Chang, Tse W. U.S. Patent 5,100,777 "Antibody matrix device and method for evaluating immune status", Priority date April 27, 1987
  9. ↑ Chang TW (March 1993). "Immunosorbent Cytometry". Biotechnology 11 (3): 291–3. doi:10.1038/nbt0393-291. PMID 7765290. 
  10. ↑ Ekins RP (1989). "Multi-analyte immunoassay". J Pharm Biomed Anal 7 (2): 155–68. doi:10.1016/0731-7085(89)80079-2. PMID 2488616. 
  11. ↑ "Multianalyte microspot immunoassay—microanalytical "compact disk" of the future". Clin. Chem. 37 (11): 1955–67. November 1991. doi:10.1016/0167-7799(94)90111-2. PMID 1934470. 
  12. ↑ Ekins RP (September 1998). "Ligand assays: from electrophoresis to miniaturized microarrays". Clin. Chem. 44 (9): 2015–30. doi:10.1093/clinchem/44.9.2015. PMID 9733000. http://www.clinchem.org/cgi/pmidlookup?view=long&pmid=9733000. 
  13. ↑ Jiang, W., Mao, Y. Q., Huang, R., Duan, C., Xi, Y., Yang, K., & Huang, R. P. (2014). "Protein expression profiling by antibody array analysis with use of dried blood spot samples on filter paper". Journal of Immunological Methods 403 (1): 79–86. doi:10.1016/j.jim.2013.11.016. PMID 24287424. 
  14. ↑ Zeng Q., Chen W. (2010). "The functional behavior of a macrophage/fibroblast co-culture model derived from normal and diabetic mice with a marine gelatin–oxidized alginate hydrogel". Biomaterials 31 (22): 5772–5781. doi:10.1016/j.biomaterials.2010.04.022. PMID 20452666. 
  15. ↑ Sohn Elliott H (2015). "Allogenic iPSC-derived RPE cell transplants induce immune response in pigs: a pilot study". Scientific Reports 5: 11791. doi:10.1038/srep11791. PMID 26138532. Bibcode: 2015NatSR...511791S. 
  16. ↑ Silzel J. W., Cercek B., Dodson C., Tsay T., Obremski R. J. (1998). "Mass-sensing, multianalyte microarray immunoassay with imaging detection". Clin. Chem. 44 (9): 2036–2043. doi:10.1093/clinchem/44.9.2036. PMID 9733002. 
  17. ↑ Mendoza L. G., McQuary P., Mongan A., Gangadharan R., Brignac S., Eggers M. (1999). "High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA)". BioTechniques 27 (4): 778–788. doi:10.2144/99274rr01. PMID 10524321. 
  18. ↑ Lueking A., Horn M., Eickhoff H., Bussow K., Lehrach H., Walter G. (1999). "Protein microarrays for gene expression and antibody screening". Anal. Biochem. 270 (1): 103–111. doi:10.1006/abio.1999.4063. PMID 10328771. 
  19. ↑ de Wildt, R. M., Mundy, C. R., Gorick, B. D., and Tomlinson, I. M. (2000) Antibody arrays for high-throughput screening of antibody-antigen interactions. Nature Biotechnol. 18, 989 –994
  20. ↑ Holt L. J., Bussow K., Walter G., Tomlinson I. M. (2000). "By-passing selection: Direct screening for antibody–antigen interactions using protein arrays". Nucleic Acids Res 28 (15): E72. doi:10.1093/nar/28.15.e72. PMID 10908365. 
  21. ↑ Zhu H., Klemic J. F., Chang S., Bertone P., Casamayor A., Klemic K. G., Smith D., Gerstein M., Reed M. A., Snyder M. (2000). "Analysis of yeast protein kinases using protein chips". Nature Genetics 26 (3): 283–289. doi:10.1038/81576. PMID 11062466. 
  22. ↑ Joos T. O., Schrenk M., Hopfl P., Kroger K., Chowdhury U., Stoll D., Schorner D., Durr M., Herick K., Rupp S., Sohn K., Hammerle H. (2000). "A microarray enzyme-linked immunosorbent assay for autoimmune diagnostics". Electrophoresis 21 (13): 2641–2650. doi:10.1002/1522-2683(20000701)21:13<2641::aid-elps2641>3.0.co;2-5. PMID 10949141. 
  23. ↑ Walter G., Bussow K., Cahill D., Lueking A., Lehrach H. (2000). "Protein arrays for gene expression and molecular interaction screening". Curr. Opin. Microbiol. 3 (3): 298–302. doi:10.1016/s1369-5274(00)00093-x. PMID 10851162. 
  24. ↑ Service R. F. (2000). "Biochemistry: Protein arrays step out of DNA's shadow". Science 289 (5485): 1673. doi:10.1126/science.289.5485.1673. PMID 11001728. 
  25. ↑ Wang Y., Wu T. R., Cai S., Welte T., Chin Y. E. (2000). "Stat1 as a Component of Tumor Necrosis Factor Alpha Receptor 1-TRADD Signaling Complex To Inhibit NF-κB Activation". Molecular and Cellular Biology 20 (13): 4505–4512. doi:10.1128/mcb.20.13.4505-4512.2000. PMID 10848577. 
  26. ↑ R.-P. Huang. (2001). Simultaneous detection of multiple proteins with an array-based enzyme-linked immunosorbant assay (ELISA) and enhanced chemiluminescence (ECL). Clin. Chem. Lab. Med. 39:209-214.
  27. ↑ "Antibody microarray-based profiling of complex specimens: systematic evaluation of labeling strategies". Proteomics 7 (11): 1786–99. 2007. doi:10.1002/pmic.200600762. PMID 17474144. 
  28. ↑ "Antibody microarray-based profiling of complex specimens: systematic evaluation of labeling strategies". Proteomics 7 (11): 1786–99. 2007. doi:10.1002/pmic.200600762. PMID 17474144. 
  29. ↑ Wingren Christer, Ingvarsson Johan, Dexlin Linda, Szul Dominika, Borrebaeck Carl A. K. (2007). "Design of recombinant antibody microarrays for complex proteome analysis: Choice of sample labeling-tag and solid support". Proteomics 7 (17): 3055–3065. doi:10.1002/pmic.200700025. PMID 17787036. 
  30. ↑ Alhamdani, MS; Schröder, C; Hoheisel, JD (Jul 6, 2009). "Oncoproteomic profiling with antibody microarrays". Genome medicine 1 (7): 68
  31. ↑ Jones V. S., Huang R. Y., Chen L. P., Chen Z. S., Fu L., Huang R. P. (2016). "Cytokines in cancer drug resistance: cues to new therapeutic strategies". Biochimica et Biophysica Acta (BBA) - Reviews on Cancer 1865 (2): 255–265. doi:10.1016/j.bbcan.2016.03.005. PMID 26993403. 
  32. ↑ Burkholder B., Burgess R.Y. R., Luo S.H., Jones V.S., Zhang W.J., Lv Z.Q., Gao C.-Y., Wang B.-L., Zhang Y.-M., Huang R.-P. (2014). "Tumor-induced perturbations of cytokines and immune cell networks". Biochimica et Biophysica Acta (BBA) - Reviews on Cancer 1845 (2): 182–201. doi:10.1016/j.bbcan.2014.01.004. PMID 24440852. 
  33. ↑ Lin Y.Expression error: Unrecognized word "et". (2013). "Peeking into the Black Box: How Cytokine Antibody Arrays Shed Light on Molecular Mechanisms of Breast Cancer Development and its Treatment". Current Proteomics 10 (4): 269–277. doi:10.2174/1570164610666131210233343. 
  34. ↑ Huang R.-P. (2007). "An array of possibilities in cancer research using cytokine antibody arrays". Expert Review of Proteomics 4 (2): 299–308. doi:10.1586/14789450.4.2.299. PMID 17425464. 
  35. ↑ "Dual-color proteomic profiling of complex samples with a microarray of 810 cancer-related antibodies". Molecular & Cellular Proteomics 9 (6): 1271–80. 2010. doi:10.1074/mcp.m900419-mcp200. PMID 20164060. 
  36. ↑ Huang R., Jiang W., Yang J., Mao Y.Q., Zhang Y., Yang W., Yang D., Burkholder B., Huang R.F., Huang R.P. (2010). "A biotin label-based antibody array for high-content profiling of protein expression". Cancer Genomics Proteomics 7 (3): 129–41. PMID 20551245. 
  37. ↑ Schwartzbaum J.Expression error: Unrecognized word "et". (2015). "Association between Prediagnostic Allergy-Related Serum Cytokines and Glioma". PLOS ONE 10 (9): e0137503. doi:10.1371/journal.pone.0137503. PMID 26352148. Bibcode: 2015PLoSO..1037503S. 
  38. ↑ Jaeger P. A.Expression error: Unrecognized word "et". (2016). "Network-driven plasma proteomics expose molecular changes in the Alzheimer's brain". Molecular Neurodegeneration 11 (1): 31. doi:10.1186/s13024-016-0105-4. PMID 27216421. 
  39. ↑ RayBiotech, Inc. Antibody Arrays. (2017). Retrieved from the RayBiotech, Inc. website http://www.raybiotech.com/antibody-array.html
  40. ↑ "Sciomics: Antibody meets microarray - scioPhospho" (in en-us). http://www.sciomics.de/services/protein-phosphorylation-analysis. 
  41. ↑ Jiang, W., Mao, Y. Q., Huang, R., Duan, C., Xi, Y., Yang, K., & Huang, R. P. (2014). "Protein expression profiling by antibody array analysis with use of dried blood spot samples on filter paper". Journal of Immunological Methods 403 (1): 79–86. doi:10.1016/j.jim.2013.11.016. PMID 24287424. 
  42. ↑ Decrop Deborah (2017). "Single-Step Imprinting of Femtoliter Microwell Arrays Allows Digital Bioassays with Attomolar Limit of Detection". ACS Applied Materials & Interfaces 9 (12): 10418–10426. doi:10.1021/acsami.6b15415. PMID 28266828. https://lirias.kuleuven.be/bitstream/123456789/576695/1/ACSAppMatInt_DecropD_Imprinting%20femtoliter%20microwell_revised3.docx. 
  43. ↑ Guo, W; Vilaplana, L; Hansson, J; Marco, P; van der Wijngaart, W (2020). "Immunoassays on thiol-ene synthetic paper generate a superior fluorescence signal". Biosensors and Bioelectronics 163: 112279. doi:10.1016/j.bios.2020.112279. PMID 32421629. 



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