Use of biosensor as a tool for double antibody sandwich immunoassay establishment: Screening of appropriate matched monoclonal antibody pair

Main Article Content

Witida Laopajon
Kantinan chuensirikulchai
Sirirat Surinkaew
Ponrut Phunpae
Chatchai Tayapiwatana
Watchara Kasinrerk
Supansa Pata


Background: For biomarker detection, double antibody sandwich platform is the most sensitive and specific immunoassay. Development of double antibody sandwich platform requires the step of appropriate antibody pair selection. However, identification of matched antibody pairs is difficult, laborious and time-consuming.

Objectives: To illustrate the application of biosensor for selecting the suitable matched pairs of antibody for double antibody sandwich platform.

Materials and methods: Double antibody sandwich immunoassay for Ag85B protein detection was selected as a model study. Biolayer interferometry biosensor was used to search matched pairs of antibody for Ag85B protein detection. To prove the obtained information, the double antibody sandwich ELISAs were employed.

Results: Biosensor results revealed that the best-matched antibody pair might be mAb clone AM85B-8B as capture antibody and mAb clone AM85B-5B or AM85B-9B as detecting antibody. The results obtained from biosensor system and ELISA were similar.

Conclusion: Biosensor can be used as a tool for effectively screening and selecting the best pairs of antibodies for any immunoassays. Biosensor method is very useful for reducing processes and time for double antibody immunoassay development.


Download data is not yet available.

Article Details

How to Cite
Laopajon, W., chuensirikulchai, K., Surinkaew, S., Phunpae, P., Tayapiwatana, C., Kasinrerk, W., & Pata, S. (2018). Use of biosensor as a tool for double antibody sandwich immunoassay establishment:: Screening of appropriate matched monoclonal antibody pair. Journal of Associated Medical Sciences, 51(2), 93-98. Retrieved from
Medical Technology


[1]. Darwish IA. Immunoassay Methods and their Applications in Pharmaceutical Analysis: Basic Methodology and Recent Advances. Int J Biomed Sci. 2006; 2(3): 217-35.

[2]. Gaskin FE, Taylor SL. Sandwich enzyme-linked immunosorbent assay (ELISA) for detection of cashew nut in foods. J Food Sci. 2011; 76(9): T218-26.

[3]. Meany DL, Sokoll LJ, Chan DW. Early Detection of Cancer: Immunoassays for Plasma Tumor Markers. Expert Opin Med Diagn. 2009; 3(6): 597-605.

[4]. Long EF. HIV screening via fourth-generation immunoassay or nucleic acid amplification test in the United States: a cost-effectiveness analysis. PLoS One. 2011; 6(11): e27625.

[5]. Ahn KC, Kim HJ, McCoy MR, Gee SJ, Hammock BD. Immunoassays and biosensors for monitoring environmental and human exposure to pyrethroid insecticides. J Agric Food Chem. 2011; 59(7): 2792-802.

[6]. Wild D, editor. The immunoassay handbook: theory and applications of ligand binding, ELISA and related techniques. 4th ed. Amsterdam: Elsevier; 2013: 7-26.

[7]. Feraudet-Tarisse C, Mazuet C, Pauillac S, Kruger M, Lacroux C, Popoff MR, et al. Highly sensitive sandwich immunoassay and immunochromatographic test for the detection of Clostridial epsilon toxin in complex matrices. PLoS One. 2017; 12(7): e0181013.

[8]. Karen L. Cox VD, Aidas Kriauciunas, Joseph Manetta, Chahrzad Montrose, and Sitta Sittampalam. Immunoassay Methods: Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2012 [cited 2018 March 3]. Available from:

[9]. Berzofsky JA, Schechter AN. The concepts of crossreactivity and specificity in immunology. Mol Immunol. 1981; 18(8): 751-63.

[10]. Porstmann T, Kiessig ST. Enzyme immunoassay techniques. An overview. J Immunol Methods. 1992; 150(1-2): 5-21.

[11]. Kiening M, Niessner R, Weller MG. Microplate-based screening methods for the efficient development of sandwich immunoassays. Analyst. 2005; 130(12): 1580-8.

[12]. Ding XX, Li XF, Deng YQ, Guo YH, Hao W, Che XY, et al. Development of a double antibody sandwich ELISA for West Nile virus detection using monoclonal antibodies against non-structural protein 1. PLoS One. 2014; 9(10): e108623.

[13]. Wang SY, Li Z, Wang XJ, Lv S, Yang Y, Zeng LQ, et al. Development of monoclonal antibody-based sandwich ELISA for detection of dextran. Monoclon Antib Immunodiagn Immunother. 2014; 33(5): 334-9.

[14]. Wilson S, Howell S. High-throughput screening in the diagnostics industry. Biochem Soc Trans. 2002; 30(4): 794-7.

[15]. Phunpae P, Chanwong S, Tayapiwatana C, Apiratmateekul N, Makeudom A, Kasinrerk W. Rapid diagnosis of tuberculosis by identification of Antigen 85 in mycobacterial culture system. Diagn Microbiol Infect Dis. 2014; 78(3): 242-8.

[16]. Laopajon W, Takheaw N, Khummuang S, Kampoun T, Cheunsirikulchai K, Kasinrerk W, et al. Antibody biosensors for the measurement and characterization of soluble CD147 molecules. Asian Pac J Allergy Immunol. 2017.

[17]. Sakamoto S, Yusakul G, Nuntawong P, Kitisripanya T, Putalun W, Miyamoto T, et al. Development of an indirect competitive immunochromatographic strip test for rapid detection and determination of anticancer drug, harringtonine. J Chromatogr B Analyt Technol Biomed Life Sci. 2017; 1048: 150-4.

[18]. Mehra B, Bhattar S, Bhalla P, Rawat D. Rapid Tests versus ELISA for Screening of HIV Infection: Our Experience from a Voluntary Counselling and Testing Facility of a Tertiary Care Centre in North India. ISRN AIDS. 2014; 2014: 296840.

[19]. Elshal MF, McCoy JP. Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA. Methods. 2006; 38(4): 317-23.

[20]. Tobias R. Fast Quantitation of Proteins and Antibodies Using the BLItz System. [Internet] California: Pall corporation; c2018 [cited 2018 3 March]; Available from:

[21]. Dayne D. BioLayer Interferometry (BLI) — How Does it Work? California: Pall corporation; 2012 [cited 2018 March 3]; Available from:

[22]. ForteBio P. Blitz technology. California: Pall corporation; 2018 [cited 2018 March 3]; Available from:

[23]. Laitinen OH, Hytonen VP, Nordlund HR, Kulomaa MS. Genetically engineered avidins and streptavidins. Cell Mol Life Sci. 2006; 63(24): 2992-3017.