Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?
Tóm tắt
Sandwich ELISA is an ideal antigen detection and quantification assay. Recently, it was used as the basic concept for high technology diagnostics. The specificity of the assay depends on the exclusion of detection cross-reactivity which arises from using two antibodies developed in different species. Since mice and rats are the common laboratory animals used to develop antigen specific antibodies. Therefore, the questions we addressed here were (1) can one use antigen-specific antibodies raised in mice and rats in the same assay to specifically detect/quantify antigens? and (2) which antibodies of the two rodents should be placed for capturing and for detection in the antigen-detection sandwich? Direct ELISA assay was used to assess for the specific reaction of the HRP-conjugated antibody to the target serum. First reaction was to compare between either conjugate anti-rat IgG (homologous) or anti-mouse IgG (heterologous) for the detection of rat sera IgG. Following the dilution factor optimization, the O.D. were 0.744±0.051 and 0.604±0.05, respectively (p= .004). The difference in mean O.D. of 0.14 reflected an unaccepted non-specific reaction. The second reaction was to compare between either conjugate anti-mouse IgG (homologous) and anti-rat IgG (heterologous) for the detection of mouse sera IgG. The recorded O.D. were 0.9414±0.14 and 0.317 ±0.141, respectively (p= .0002). The improved difference in mean O.D. of 0.624 reflecting a minimized cross-reaction. Our results suggest that it is possible to use both Swiss albino mice and albino rats in a single sandwich ELISA, given that the captured antibody species to be from the Swiss albino mice and the detection antibody to be from the albino rat. The described working details are limited to the source of the antibodies used in the study. However, the approach stresses on the importance of such optimization steps before making any interpretations based on the antigen detection. To our knowledge, this study is the first to cover the optimal order of the capturing and the detection antibodies in a sandwich ELISA assay. In addition to addressing the possible interfering cross-reactivity that result from using mouse and rat serum antibodies in a single assay.
Tài liệu tham khảo
Karen L, Viswanath D, Aidas K et al (2004) (2012, updated in 2019) Assay Guidance Manual. In: Sittampalam GS, Grossman A, Brimacombe K et al (eds) Immunoassay Methods. Eli Lilly & Company and the National Center for Advancing Translational Sciences Available from: https://www.ncbi.nlm.nih.gov/books/NBK92434/
Weiland G (1978) The enzyme-linked immunosorbent assay (ELISA)--a new serodiagnostic method for the detection of parasitic infections (author’s transl). MMW, Munchener Medizinische Wochenschrift 120(44):1457–1460
Tenda K, van Gerven B, Arts R et al (2018) Paper-based antibody detection devices using bioluminescent BRET-switching sensor proteins. Angew Chem Int Ed 57(47):15369–15373. https://doi.org/10.1002/anie.201808070
Wu J, Dong M, Rigatto C, Liu Y, Lin F (2018) Lab-on-chip technology for chronic disease diagnosis. NPJ Digit Med 1(1):7. https://doi.org/10.1038/s41746-017-0014-0
Bayat M, Khabiri A, Hemati B (2018) Development of IgY-based sandwich ELISA as a robust tool for rapid detection and discrimination of toxigenic Vibrio cholerae. Can J Infect Dis Med Microbiol 2018:4032531–4032539. https://doi.org/10.1155/2018/4032531
Zhang WW, Ghosh AK, Mohamath R, Whittle J, Picone A, Lypaczewski P, Ndao M, Howard RF, Das P, Reed SG, Matlashewski G (2018) Development of a sandwich ELISA to detect Leishmania 40S ribosomal protein S12 antigen from blood samples of visceral Leishmaniasis patients. BMC Infect Dis 18(1):500. https://doi.org/10.1186/s12879-018-3420-2
Akin H, Akalin H, Budak F, Ener B, Ocakoğlu G, Gürcüoğlu E, Göral G, Oral HB (2015) Alterations of serum cytokine levels and their relation with inflammatory markers in candidemia. Med Mycol 53(3):258–268. https://doi.org/10.1093/mmy/myu084
Ming F, Cheng Y, Ren C, Suolang S, Zhou H (2018) Development of A DAS-ELISA for detection of H9N2 avian influenza virus. JVirol Methods 263:38–43. https://doi.org/10.1016/j.jviromet.2018.10.014
Ahmad MH, Shakeel MT, Al-Shahwan IM (2018) Insights into the incidence of watermelon chlorotic stunt virus causing yellowing disease of watermelon in western and southwestern regions of Saudi Arabia. Plant Pathol J 34(5):426–434. https://doi.org/10.5423/PPJ.OA.04.2018.0059
Dinçel E, Özkan Y, Şüküroğlu MA (2017) Evaluation of tryptophan/kynurenine pathway relevance with immune system biomarkers of low energy trauma hip fractures in osteoporotic patients. Arch Rheumatol 32(3):203–208. https://doi.org/10.5606/ArchRheumatol.2017.6216
Bontekoe J, Lee J, Bansal V, Syed M, Hoppensteadt D, Maia P, Walborn A, Liles J, Brailovsky E, Fareed J (2018) Biomarker profiling in stage 5 chronic kidney disease identifies the relationship between angiopoietin-2 and atrial fibrillation. Clin Appl Thromb Hemost 24(9):269S–276S. https://doi.org/10.1177/1076029618808909
Tyler E, Jane S, Mark AR et al (2018) Semen AMACR protein as a novel method for detecting prostate cancer. Urol Oncol 36(12):532.e1–532.e7. https://doi.org/10.1016/j.urolonc.2018.09.010
Kunireddy N, Jacob R, Khan SA, Yadagiri B, Sai Baba KSS, Rajendra Vara Prasad I, Mohan IK (2018) Hepcidin and ferritin: important mediators in inflammation associated anemia in systemic lupus erythematosus patients. Indian J Clin Biochem 33(4):406–413. https://doi.org/10.1007/s12291-017-0702-1
Yamamoto-Imoto H, Zamolodchikov D, Chen ZL, Bourne SL, Rizvi S, Singh P, Norris EH, Weis-Garcia F, Strickland S (2018) A novel detection method of cleaved plasma high-molecular-weight kininogen reveals its correlation with Alzheimer’s pathology and cognitive impairment. Alzheimers Dement 10(1):480–489. https://doi.org/10.1016/j.dadm.2018.06.008
Toinon A, Fontaine C, Thion L, Gajewska B, Carpick B, Nougarede N, Uhlrich S (2018) Host cell protein testing strategy for hepatitis B antigen in hexavalent vaccine - towards a general testing strategy for recombinant vaccines. Biologicals. 54:1–7. https://doi.org/10.1016/j.biologicals.2018.05.006
Dain K, Hyeseon Y, Sangkyu K et al (2018) Generation of a human monoclonal antibody to cross-reactive material 197 (CRM197) and development of a sandwich ELISA for CRM197 conjugate vaccines. J Microbiol Biotechnol 28(12):2113–2120
Farshidi N, Moghaddam M, Yaghoubi V, Ayati SH, Varasteh AR, Sankian M (2019) Type IV chitinase quantification in four different grape cultivars (Vitis vinifera) in northeast of Iran by an indirect sandwich enzyme-linked immunosorbent assay. J Immunoass Immunochem 40(2):139–148. https://doi.org/10.1080/15321819.2018.1535439
Akbari NS, Khalilzadeh B, Samadi Pakchin P et al (2018) Highly sensitive and reliable detection of CA15-3 in patient plasma with electrochemical biosensor labeled with magnetic beads. Biosens Bioelectron 122:8–15. https://doi.org/10.1016/j.bios.2018.08.047
Weil GJ, Lammie PJ, Weiss N (1997) The ICT filariasis test: a rapid-format antigen test for diagnosis of bancroftian filariasis. Parasitol Today 13(10):401–404. https://doi.org/10.1016/s0169-4758(97)01130-7
Chesnais CB, Missamou F, Pion SD et al (2013) Semi-quantitative scoring of an immunochromatographic test for circulating filarial antigen. Am J Trop Med Hyg 89(5):916–918. https://doi.org/10.4269/ajtmh.13-0245
Chesnais CB, Vlaminck J, Kunyu-Shako B, Pion SD, Awaca-Uvon NP, Weil GJ, Mumba D, Boussinesq M (2016) Measurement of circulating filarial antigen levels in human blood with a point-of-care test strip and a portable spectrodensitometer. Am J Trop Med Hyg 94(6):1324–1329. https://doi.org/10.4269/ajtmh.15-0916
Jianwen H (2013) Practical guide to ELISA development. In: The Immunoassay Handbook Theory and applications of ligand binding, ELISA and related techniques, 4th edn. Elsevier Ltd
Shani B, Hector M, Osnat P et al (2009) Rodent phylogeny revised: analysis of six nuclear genes from all major rodent clades. BMC Evol Biol 9(1):71. https://doi.org/10.1186/1471-2148-9-71
de Winter JCF (2013) Using the Student’s-t-test with extremely small sample sizes. Pract Assess Res Eval 18:10. https://doi.org/10.7275/e4r6-dj05
Nahm M, Der-Balian GP, Venturini D et al (1980) Antigenic similarities of rat and mouse IgG subclasses associated with anti-carbohydrate specificities. Immunogenetics. 11(2):199–203. https://doi.org/10.1007/bf01567785