Cytokine detection and simultaneous assessment of rheumatoid factor interference in human serum and synovial fluid using high-sensitivity protein arrays on plasmonic gold chips

BMC Biotechnology

Manfè Valentina²  Fleckner Jan¹  Nørby Lisby Peder¹  Zhang Bo²  Dai Hongjie²  Keller Pernille¹ 

[1] Department of PharmacoGenetics, Biopharmaceutical Research Unit, Novo Nordisk A/S, Novo Nordisk Park 1, Maaloev 2760, Denmark

[2] Department of Chemistry, Stanford University, San Francisco, CA, USA

关键词: Fluorescence enhancement;    Plasmonic gold;    Rheumatoid factor interference;    Osteoarthritis;    Rheumatoid arthritis;    IL-20;    Cytokines;    Protein microarray;   

Others  :  1222684
DOI  :  10.1186/s12896-015-0186-0

 received in 2015-01-19, accepted in 2015-07-24,  发布年份 2015

【 摘 要 】

Background

Fluorescence-enhancing microarray on plasmonic gold film is an attractive alternative to traditional enzyme-linked immunosorbent assay (ELISA) for cytokine detection because of the increased sensitivity. The assay chemistry is similar to an ELISA sandwich assay, but owing to the gold substrate, cytokine measurements are 10 to 100 times more sensitive and can be multiplexed. Plasmonic protein microarrays are, as other immunoassays, affected by the presence of heterophilic antibodies and rheumatoid factor may lead to analytical errors with serious implications for patient care. Here, we present a plasmonic gold substrate protein microarray for high-sensitivity detection of cytokines with simultaneous assessment of rheumatoid factor interference on a single chip.

Results

Paired serum and synovial fluid samples from patients with rheumatoid arthritis (n = 18), osteoarthritis (n = 9) or healthy controls (n = 10) were arrayed on near-infrared fluorescence enhancing plasmonic gold chips spotted with cytokine-specific capture antibody and isotype control antibody. Possible rheumatoid factor interference was visualised by a non-specific signal from the isotype control antibody, and pre-treatment of samples with heat-aggregated animal IgG eliminated this background contamination. The platform was optimised using the cytokine IL-20. The protein microarray platform allowed for the detection of human IL-20 at levels <1 pg/ml with reliable IL-20 quantification over a 5-log dynamic range. Samples for which rheumatoid factor caused artefacts were identified and a method for eliminating rheumatoid factor interference was developed and validated. IL-20 protein levels were significantly higher in synovial fluid samples from patients with rheumatoid arthritis compared to osteoarthritis (p < 0.001), while serum levels of IL-20 did not differ between patients with rheumatoid arthritis, osteoarthritis or healthy controls.

Conclusion

Using novel plasmonic gold chips, we developed a highly sensitive and accurate assay platform to detect lowly expressed cytokines in biological fluids, allowing for the elimination of rheumatoid factor interference in as little as 5 μl sample volume. The detection limit was below 1 pg/ml for IL-20 and linearity was achieved over a 5-log dynamic range. This technology is highly advantageous for cytokines where sensitivity or sample volume is critical or where assessment of rheumatoid factor interference needs addressed and eliminated.

【 授权许可】

   
2015 Valentina et al.

【 图 表 】

Fig 1.

Fig 2.

Fig 1.

Fig 2.

【 参考文献 】

• [1]Feldmann M. Many cytokines are very useful therapeutic targets in disease. J Clin Invest. 2008; 118:3533-3536.
• [2]Cox KL, Devanarayan V, Kriauciunas A, Manetta J, Montrose C, Sittampalam S: Immunoassay Methods. 2004.
• [3]Selvarajah S, Negm OH, Hamed MR, Tubby C, Todd I, Tighe PJ, Harrison T, Fairclough LC. Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera. Mediators Inflamm. 2014; 2014:820304.
• [4]Tabakman SM, Lau L, Robinson JT, Price J, Sherlock SP, Wang H, Zhang B, Chen Z, Tangsombatvisit S, Jarrell JA, Utz PJ, Dai H. Plasmonic substrates for multiplexed protein microarrays with femtomolar sensitivity and broad dynamic range. Nat Commun. 2011; 2:466.
• [5]Zhang B, Jarrell JA, Price JV, Tabakman SM, Li Y, Gong M, Hong G, Feng J, Utz PJ, Dai H. An integrated peptide-antigen microarray on plasmonic gold films for sensitive human antibody profiling. PLoS One. 2013; 8:e71043.
• [6]Zhang B, Kumar RB, Dai H, Feldman BJ. A plasmonic chip for biomarker discovery and diagnosis of type 1 diabetes. Nat Med. 2014; 20:948-953.
• [7]Zhang B, Price J, Hong G, Tabakman SM, Wang H, Jarrell JA, Hong G, Utz PJ, Dai H. Multiplexed cytokine detection on plasmonic gold substrates with enhanced near-infrared fluorescence. Nano Research 2013.
• [8]Leng RX, Pan HF, Tao JH, Ye DQ. IL-19, IL-20 and IL-24: potential therapeutic targets for autoimmune diseases. Expert Opin Ther Targets. 2011; 15:119-126.
• [9]Otkjaer K, Kragballe K, Funding AT, Clausen JT, Noerby PL, Steiniche T, Iversen L. The dynamics of gene expression of interleukin-19 and interleukin-20 and their receptors in psoriasis. Br J Dermatol. 2005; 153:911-918.
• [10]Hsu YH, Li HH, Hsieh MY, Liu MF, Huang KY, Chin LS, Chen PC, Cheng HH, Chang MS. Function of interleukin-20 as a proinflammatory molecule in rheumatoid and experimental arthritis. Arthritis Rheum. 2006; 54:2722-2733.
• [11]Hsing CH, Cheng HC, Hsu YH, Chan CH, Yeh CH, Li CF, Chang MS. Upregulated IL-19 in breast cancer promotes tumor progression and affects clinical outcome. Clin Cancer Res. 2012; 18:713-725.
• [12]Wei CC, Hsu YH, Li HH, Wang YC, Hsieh MY, Chen WY, Hsing CH, Chang MS. IL-20: biological functions and clinical implications. J Biomed Sci. 2006; 13:601-612.
• [13]Lee SJ, Cho SC, Lee EJ, Kim S, Lee SB, Lim JH, Choi YH, Kim WJ, Moon SK. Interleukin-20 promotes migration of bladder cancer cells through extracellular signal-regulated kinase (ERK)-mediated MMP-9 protein expression leading to nuclear factor (NF-kappaB) activation by inducing the up-regulation of p21(WAF1) protein expression. J Biol Chem. 2013; 288:5539-5552.
• [14]Aydogdu E, Pamuk ON, Donmez S, Pamuk GE. Decreased interleukin-20 level in patients with systemic sclerosis: are they related with angiogenesis? Clin Rheumatol. 2013; 32:1599-1603.
• [15]Kragstrup TW, Otkjaer K, Holm C, Jorgensen A, Hokland M, Iversen L, Deleuran B. The expression of IL-20 and IL-24 and their shared receptors are increased in rheumatoid arthritis and spondyloarthropathy. Cytokine. 2008; 41:16-23.
• [16]Todd DJ, Knowlton N, Amato M, Frank MB, Schur PH, Izmailova ES, Roubenoff R, Shadick NA, Weinblatt ME, Centola M, Lee DM. Erroneous augmentation of multiplex assay measurements in patients with rheumatoid arthritis due to heterophilic binding by serum rheumatoid factor. Arthritis Rheum. 2011; 63:894-903.
• [17]Gonzalez A, Icen M, Kremers HM, Crowson CS, Davis JM, Therneau TM, Roger VL, Gabriel SE. Mortality trends in rheumatoid arthritis: the role of rheumatoid factor. J Rheumatol. 2008; 35:1009-1014.
• [18]Moore TL, Dorner RW. Rheumatoid factors. Clin Biochem. 1993; 26:75-84.
• [19]Ward G, McKinnon L, Badrick T, Hickman PE. Heterophilic antibodies remain a problem for the immunoassay laboratory. Am J Clin Pathol. 1997; 108:417-421.
• [20]Kragstrup TW, Vorup-Jensen T, Deleuran B, Hvid M. A simple set of validation steps identifies and removes false results in a sandwich enzyme-linked immunosorbent assay caused by anti-animal IgG antibodies in plasma from arthritis patients. Springerplus. 2013; 2:263. BioMed Central Full Text
• [21]Tate J, Ward G. Interferences in immunoassay. Clin Biochem Rev. 2004; 25:105-120.
• [22]Scrivo R, Conigliaro P, Riccieri V, Di FM, Alessandri C, Spadaro A, Perricone R, Valesini G. Distribution of IL-10 family cytokines in serum and synovial fluid of patients with inflammatory arthritis reveals different contribution to systemic and joint inflammation. Clin Exp Immunol. 2015; 179(2):300-8.
• [23]Sturgeon CM, Viljoen A. Analytical error and interference in immunoassay: minimizing risk. Ann Clin Biochem. 2011; 48:418-432.
• [24]Tiwari G, Tiwari R. Bioanalytical method validation: An updated review. Pharm Methods. 2010; 1:25-38.