Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b

Xi Qin; Maoqin Duan; Dening Pei; Jian Lin; Lan Wang; Peng Zhou; Wenrong Yao; Ying Guo; Xiang Li; Lei Tao; Youxue Ding; Lan Liu; Yong Zhou; Chuncui Jia; Chunming Rao; Junzhi Wang

doi:10.1016/j.jpha.2021.07.003

Volume 12 Issue 2

May 2022

Turn off MathJax

Article Contents

Article Navigation > Journal of Pharmaceutical Analysis > 2022 > 12(2): 308-316

Xi Qin, Maoqin Duan, Dening Pei, Jian Lin, Lan Wang, Peng Zhou, Wenrong Yao, Ying Guo, Xiang Li, Lei Tao, Youxue Ding, Lan Liu, Yong Zhou, Chuncui Jia, Chunming Rao, Junzhi Wang. Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b[J]. Journal of Pharmaceutical Analysis, 2022, 12(2): 308-316. doi: 10.1016/j.jpha.2021.07.003

Citation:

Xi Qin, Maoqin Duan, Dening Pei, Jian Lin, Lan Wang, Peng Zhou, Wenrong Yao, Ying Guo, Xiang Li, Lei Tao, Youxue Ding, Lan Liu, Yong Zhou, Chuncui Jia, Chunming Rao, Junzhi Wang. Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b[J]. Journal of Pharmaceutical Analysis, 2022, 12(2): 308-316. doi: 10.1016/j.jpha.2021.07.003

Citation:

Xi Qin, Maoqin Duan, Dening Pei, Jian Lin, Lan Wang, Peng Zhou, Wenrong Yao, Ying Guo, Xiang Li, Lei Tao, Youxue Ding, Lan Liu, Yong Zhou, Chuncui Jia, Chunming Rao, Junzhi Wang. Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b[J]. Journal of Pharmaceutical Analysis, 2022, 12(2): 308-316. doi: 10.1016/j.jpha.2021.07.003

PDF( 1845 KB)

Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b

doi: 10.1016/j.jpha.2021.07.003

a. National Institutes for Food and Drug Control, Beijing, 100050, China;
b. Synthetic and Functional Biomolecules Center, Peking University, Beijing, 100871, China;
c. WHO Collaboration Centre for Biologicals Standardization and Evaluation, Beijing, 100050, China

Funds:

Financial support was provided by the National Science and Technology Major Project (Grant No.: 2015ZX09501008). The fund agency did not participate in the study's design, data collection and analysis, decision to publish, and preparation of the manuscript. We would like to thank Dr. Weijun Ma from the Shanghai Institutes for Biological Sciences for the provision of the vector Rphen1 and helper phages.

Received Date: Jul. 05, 2020
Accepted Date: Jul. 05, 2021
Rev Recd Date: May 25, 2021
Publish Date: Jul. 08, 2021

Abstract

Abstract

Recombinant human interferon α2b (rhIFNα2b) is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C. The current identification test for rhIFNα2b is complex. In this study, an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b. RhIFNα2b was used to immunize an alpaca, which established a phage nanobody library. After five steps of enrichment, the nanobody I22, which specifically bound rhIFNα2b, was isolated and inserted into the prokaryotic expression vector pET28a. After subsequent purification, the physicochemical properties of the nanobody were determined. A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody. To develop a rapid test, the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips. The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL. The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products. The principle of this novel assay is generally applicable for the rapid testing of other commercial products, with a great potential for routine use in detecting counterfeit recombinant protein products.
- Recombinant human interferon α2b,
- Nanobody,
- Phage display,
- Screening,
- Rapid test

FullText(HTML)

References(34)

References

A. Isaacs, J. Lenmann, R.C. Valentine, Virus interference. II. Some properties of interferon, Proc. R. Lond. B. Biol. Sci. 147 (1957) 268-273

A. Isaacs, J. Lenmann, R.C. Valentine, Virus interference. I. The interferon, Proc. R. Lond. B. Biol. Sci. 147 (1957) 258-267

J.Z. Wang, Research, Development and Quality Control of Biopharmaceuticals, 3rd ed., Science Press, Beijing, 2018, pp. 467-483

Chinese Pharmacopoeia Commission, Pharmacopoeia of the People's Republic of China (Part III), 2020 edition, China Medical Science Press, Beijing, 2020, pp. 309-320

C. Hamers-Casterman, T. Atarhouch, S. Muyldermans, et al., Naturally occurring antibodies devoid of light chains, Nature. 363 (1993) 446-448

C. Li, Z. Tang, Z. Hu, et al., Natural Single-Domain Antibody-Nanobody: A Novel Concept in the Antibody Field, J. Biomed. Nanotechnol. 14 (2018) 1-19

P. Bannas, J. Hambach, F. Koch-Nolte, Nanobodies and Nanobody-Based Human Heavy Chain Antibodies as Antitumor Therapeutics, Front. Immunol. 8 (2017), 1603

S. Muyldermans, T.N. Baral, V.C. Retamozzo, et al., Camelid immunoglobulins and nanobody technology, Vet. Immunol. Immunopathol. 128 (2009) 178-183

L.S. Mitchell, L.J. Colwell, Comparative analysis of nanobody sequence and structure data, Proteins. 86 (2018) 697-706

M. Yamagata, J.R. Sanes, Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical reagents, Proc. Natl. Acad. Sci. USA. 115 (2018) 2126-2131

M. Dumoulin, K. Conrath, A. Van Meirhaeghe, et al., Single-domain antibody fragments with high conformational stability, Protein Sci. 11 (2002) 500-515

S. Muyldermans, Nanobodies: natural single-domain antibodies, Annu. Rev. Biochem. 82 (2013) 775-797

R.H. Van der Linden, L.G. Frenken, B. de Geus, et al., Comparison of physical chemical properties of llama VHH ant ibody fragments and mouse monoclonal antibodies, Biochem. Biophys. Acta. 1431 (1999) 37-46

B. Stijlemans, K. Conrath, V. Cortez-Retamozo, et al., Efficient targeting of conserved cryptic epitopes of infectious agents by single-domain antibodies. African trypanosomes as a paradigm, J. Biol. Chem. 279 (2004) 1256-1261

P.D. Skottrup, Structural insights into a high affinity nanobody: antigen complex by homology modelling, J. Mol. Graph. Model. 76 (2017) 305-312

V. Cortez-Retamozo, N. Backmann, P.D. Senter, et al., Efficient cancer therapy with a nanobody-based conjugate, Cancer Res. 64 (2004) 2853-2857

V. Cortez-Retamozo, M. Lauwereys, Gh. G. Hassanzadeh, et al., Efficient tumor targeting by single-domain antibody fragments of camels, Int. J. Cancer. 98 (2002) 456-462

D. Pan, G.H. Li, H.Z. Hu, et al., Direct Immunoassay for Facile and Sensitive Detection of Small Molecule Aflatoxin B1 based on Nanobody, Chemistry. 24 (2018) 9869-9876

D.R. Maass, J. Sepulveda, A. Pernthaner, et al., Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs), J. Immunol. Methods. 324 (2007) 13-25

L. Aujame, F. Geoffroy, R. Sodoyer, High affinity human antibodies by phage display, Hum. Antibodies. 8 (1997) 155-168

M. Zhu, X. Gong, Y. Hu, et al., Streptavidin-biotin-based directional double Nanobody sandwich ELISA for clinical rapid and sensitive detection of influenza H5N1, J. Transl. Med. 12 (2014), 352

N. Kobayashi, T. Karibe, J. Goto, Dissociation-independent selection of high-affinity anti-hapten phage antibodies using cleavable biotin-conjugatens, Anal. Biochem. 347 (2005) 287-296

A. Pini, C. Ricci, L. Bracci, Phage display and colony filter screening for high-throughput selection of antibody libraries, Comb. Chem. High Throughput Screen. 5 (2002) 503-510

R. Baghban, S.L. Gargari, M. Rajabibazl, et al., Camelid-derived heavy-chain nanobody against Clostridium botulinum neurotoxin E in Pichia pastoris, Biotechnol. Appl. Biochem. 63 (2016) 200-205

C. Hemmer, S. Djennane, L. Ackerer, et al., Nanobody-mediated resistance to Grapevine fanleaf virus in plants, Plant Biotechnol. J. 16 (2018) 660-671

M. Behdani, S. Zeinali, M. Karimipour, et al., Development of VEGFR2-specific Nanobody Pseudomonas exotoxin A conjugated to provide efficient inhibition of tumor cell growth, N. Biotechnol. 30 (2013) 205-209

P. Schotte, I. Dewerte, M. De Groeve, et al., Pichia astoris Mut (S) strains are prone to misincorporation of O-methyl-L-homoserine at methionine residues when methanol is used as the sole carbon source, Microb. Cell Fact. 15 (2016), 98

J.L. Liu, D. Zabetakis, E.R. Goldman, et al., Selection and characterization of single domain antibodies against human CD20, Mol. Immunol. 78 (2016) 146-154

R.C. Stevens, Design of high-throughput methods of protein production for structure biology, Structure 8 (2000) R177-R185

D.S. Waugh, Making the most of affinity tags, Trends. Biotechnol. 23 (2005) 316-320

K. Terpe, Overview of tag protein fusion: from molecular and biochemical fundamentals to commercial systems, Appl. Microbiol. Biotechnol. 60 (2003) 523-533

E. Soler, L.M. Houdebine, Preparation of recombinant vaccines, Biotechnol. Annu. Rev. 13 (2007) 65-94

P. Kunz, K. Zinner, N. Mucke, et al., The structural basis of nanobody unfolding reversibility and thermos-resistance, Sci. Rep. 8 (2018), 7934

Y. Wang, Z. Fan, L. Shao, et al., Nanobody-derived nanobiotechnology tool kits for diverse biomedical and biotechnology applications, Int. J. Nanomedicine. 11 (2016) 3287-3303

Relative Articles

Supplements(0)

Cited By

Proportional views