Abstract
Recombinant antibody fragments like single chain variable fragments (scFvs) represent an attractive yet powerful alternative to immunoglobulins and hold great potential in the development of clinical diagnostic/therapeutic reagents. Structurally, scFvs are the smallest antibody fragments capable of retaining the antigen-binding capacity of whole antibodies and are composed of an immunoglobulin (Ig) variable light (VL) and variable heavy (VH) chain joined by a flexible polypeptide linker. In the present study, we constructed a scFv against bovine IgA from a hybridoma cell line IL-A71 that secretes a monoclonal antibody against bovine IgA using recombinant DNA technology. The scFv was expressed in Escherichia coli and purified using immobilized metal affinity chromatography (IMAC). The binding activity and specificity of the scFv was established by its non-reactivity toward other classes of immunoglobulins as determined by enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. Kinetic measurement of the scFv indicated that the recombinant antibody fragment had an affinity in picomolar range toward purified IgA. Furthermore, the scFv was used to develop a sensitive ELISA for the detection of foot and mouth disease virus (FMDV) carrier animals.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1. K.B. Renegar P.A. Small 1991 Passive transfer of local immunity to influenza virus infection by IgA antibody J Immunol 146 1972 1978.
-
2. L. Winner J. Mack R. Weltzin J.J. Mekalanos J.P. Kraehenbuhl 1991 New model for analysis of mucosal immunity: intestinal secretion of specific monoclonal immunoglobulin A from hybridoma tumors protects against Vibrio cholerae infection Infect Immun 59 977 982.
-
3. H. Stubbe J. Berdoz J.P. Kraehenbuhl B. Corthésy 2000 Polymeric IgA is superior to monomeric IgA and IgG carrying the same variable domain in preventing Clostridium difficile toxin A damaging of T84 monolayers J Immunol 164 1952 1960.
-
4. V. Friman F. Nowrouzian I. Adlerberth A.E. Wold 2002 Increased frequency of intestinal Escherichia coli carrying genes for S fimbriae and haemolysin in IgA deficient individuals Microb Pathog 32 35 42.
-
5. P.R. Burnett T.C. VanCott V.R. Polonis R.R. Redfield D.L. Birx 1994 Serum IgA-mediated neutralization of HIV type 1 J Immunol 152 4642 4648.
-
6. M.W. Russell M. Kilian 2005 Biological activities of IgA M. J B. J L. ME M. L S. W Mucosal Immunology Academic Press San Diego 267 289.
-
7. R. Orlandi D.H. Gussow P.T. Jones G. Winter 1989 Cloning immunoglobulin variable domains for expression by the polymerase chain reaction Proc Natl Acad Sci USA 86 3833 3837.
-
8. J. McCafferty A.D. Griffiths G. Winter D.J. Chiswell 1990 Phage antibodies: filamentous phage displaying antibody variable domains Nature 348 552 554.
-
9. J. Brumester A. Pluckthun 2001 Construction of scFv fragments from hybridoma or spleen cells by PCR assembly K. R D. S Antibody Engineering Springer-Verlag Berlin, Germany 19 40.
-
10. J.S. Huston D. Levinson M. Mudgett-Hunter M.S. Tai J. Novotny 1988 Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli Proc Natl Acad Sci USA 85 5879 5883.
-
11. R.E. Bird K.D. Hardman J.W. Jacobson S. Johnson B.M. Kaufman 1988 Single-chain antigen-binding proteins Science 242 423 426.
-
12. J.H. Condra V.V. Sardana J.E. Tomassini A.J. Schlabach M.E. Davies 1990 Bacterial expression of antibody fragments that block human rhinovirus infection of cultured cells J Biol Chem 265 2292 2295.
-
13. L.H. Chen Q. Huang L. Wan L.Y. Zeng S.F. Li 2006 Expression, purification, and in vitro refolding of a humanized single-chain Fv antibody against human CTLA4 (CD152) Protein Expr Purif 46 495 502.
-
14. S. Padiolleau-Lefevre C. Alexandrenne F. Dkhissi G. Clement S. Essono 2007 Expression and detection strategies for an scFv fragment retaining the same high affinity than Fab and whole antibody: Implications for therapeutic use in prion disease Mol Immunol 44 1888 1896.
-
15. P. Holliger P.J. Hudson 2005 Engineered antibody fragments and the rise of single domains Nat Biotechnol 23 1126 1136.
-
16. P.J. Carter 2006 Potent antibody therapeutics by design Nat Rev Immunol 6 343 357.
-
17. D. Filpula 2007 Antibody engineering and modification technologies Biomol Eng 24 201 215.
-
18. B. Luginbuhl Z. Kanyo R.M. Jones R.J. Fletterick S.B. Prusiner 2006 Directed evolution of an anti-prion protein scFv fragment to an affinity of 1 pM and its structural interpretation J Mol Biol 363 75 97.
-
19. M. Amadori B. Hass A. Moos I. Zerbini 2002 IgA response of cattle FMDV infection in probang and saliva samples EU FMD Ras. Gr. Borovets 88 106.
-
20. J.S. Salt G. Mulcahy R.P. Kitching 1996 Isotype-specific antibody responses to foot-and-mouth disease virus in sera and secretions of carrier and non-carrier cattle Epidemiol Infect 117 349 360.
-
21. I.L. Archetti M. Amadori A. Donn J. Salt E. Lodetti 1995 Detection of foot-and-mouth disease virus-infected cattle by assessment of antibody response in oropharyngeal fluids J Clin Microbiol 33 79 84.
-
22. J.D. Marks H.R. Hoogenboom A.D. Griffiths G. Winter 1992 Molecular evolution of proteins on filamentous phage J Biol Chem 267 16007 16010.
-
23. U.K. Laemmli 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature 227 5259 680 685.
-
24. J. Mestecky M.W. Russell C.O. Elson 1999 Intestinal IgA: novel views on its function in the defence of the largest mucosal surface Gut 44 2 5.
-
25. M.B. Mazanec C.S. Kaetzel M.E. Lamm D. Fletcher J.G. Nedrud 1992 Intracellular neutralization of virus by immunoglobulin A antibodies Proc Natl Acad Sci USA 89 6901 6905.
-
26. M.B. Mazanec J.G. Nedrud C.S. Kaetzel M.E. Lamm 1993 A three-tiered view of the role of IgA in mucosal defense Immunol Today 14 430 435.
-
27. M.B. Mazanec C.L. Coudret D.R. Fletcher 1995 Intracellular neutralization of influenza virus by immunoglobulin A antihemagglutinin monoclonal antibodies J Virol 69 1339 1343.
-
28. C.S. Kaetzel J.K. Robinson K.R. Chintalacharuvu J.P. Vaerman M.E. Lam 1991 The polymeric immunoglobulin receptor (secretory component) mediates transport of immune complexes across epithelial cells: a local defense function for IgA Proc Natl Acad Sci USA 88 8796 8800.
-
29. M.J. Feldhaus R.W. Siegel L.K. Opresko 2003 Flow cytometric isolation of human antibodies from a nonimmune Saccharomyces cerevisiae surface display library Nat Biotechnol 21 2 163 170.
-
30. J. Hanes L. Jermutus S. Weber-Bornhauser H.R. Bosshard A. Plückthun 1998 Ribosome display efficiently selects and evolves high-affinity antibodies in vitro from immune libraries Proceedings of the National Academy of Sciences of the United States of America 95 24 4130 14135.
-
31. R. Orlandi D.H. Güssow P.T. Jones G. Winter 1989 Cloning immunoglobulin variable domains for expression by the polymerase chain reaction Proc Natl Acad Sci USA 86 10 3833 3837.
-
32. J. McCafferty A.D. Griffiths G. Winter D.J. Chiswell 1990 Phage antibodies: Filamentous phage displaying antibody variable domains Nature 348 552 554.
-
33. L.K. Gilliland N.A. Norris H. Marquardt T.T. Tsu M.S. Hayden 1996 Rapid and reliable cloning of antibody variable regions and generation of recombinant single chain antibody fragments Tissue Antigens 47 1 1 20.
-
34. M. Ali K. Hitomi H. Nakano 2006 Generation of monoclonal antibodies using simplified single cell reverse transcriptionpolymerase chain reaction and cell free protein synthesis J Biosci Bioeng 101 284 286.
-
35. J.F. Adam J.D. Muller M. Yu L.-F. Wang G.H. Hans 2007 Production and application of recombinant antibodies to FMDV non structural protein 3ABC J Immunol Methods 321 142 151.
-
36. S.C. Cheung B. Dietzschold H. Koprowski A.L. Notkins R.F. Rando 1992 A recombinant human Fab expressed in Escherichia coli neutralizes rabies virus J Virol 66 6714 6720.
Please, download the file from HERE.
Dr. Dunay, Ildiko Rita
Magdeburg, Germany
E-mail: ildikodunay@gmail.com
Indexing and Abstracting Services:
- PubMed Central
- Scopus
- ESCI
- CABI
- CABELLS Journalytics
| 2021 | |
|---|---|
| Web of Science | |
| Total Cites WoS |
790 |
| Journal Impact Factor | not applicable |
| Rank by Impact Factor | not applicable |
| Impact Factor without Journal Self Cites |
not applicable |
| 5 Year Impact Factor |
not applicable |
| Journal Citation Indicator | 0,64 |
| Rank by Journal Citation Indicator | Microbiology 81/157 |
| Scimago | |
| Scimago H-index |
not indexed |
| Scimago Journal Rank |
not indexed |
| Scimago Quartile Score | not indexed |
| Scopus | |
| Scopus Cite Score |
not indexed |
| Scopus CIte Score Rank |
not indexed |
| Scopus SNIP |
not indexed |
| 2020 | |
|---|---|
| CrossRef Documents | 23 |
| WoS Cites | 708 |
| Wos H-index | 27 |
| Days from submission to acceptance | 219 |
| Days from acceptance to publication | 176 |
| Acceptance Rate | 70% |
| 2019 | |
|---|---|
| WoS Cites |
558 |
| CrossRef Documents |
24 |
| Acceptance Rate |
92% |
| European Journal of Microbiology and Immunology | |
|---|---|
| Publication Model | Gold Open Access |
| Submission Fee | none |
| Article Processing Charge | 600 EUR/article |
| Regional discounts on country of the funding agency | World Bank Lower-middle-income economies: 50% World Bank Low-income economies: 100% |
| Further Discounts | Editorial Board / Advisory Board members: 50% Corresponding authors, affiliated to an EISZ member institution subscribing to the journal package of Akadémiai Kiadó: 100% |
| Subscription Information | Gold Open Access |
| Purchase per Title | |
| European Journal of Microbiology and Immunology | |
|---|---|
| Language | English |
| Size | A4 |
| Year of Foundation |
2011 |
| Volumes per Year |
1 |
| Issues per Year |
4 |
| Founder | Akadémiai Kiadó |
| Founder's Address |
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245. |
| Publisher | Akadémiai Kiadó |
| Publisher's Address |
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245. |
| Responsible Publisher |
Chief Executive Officer, Akadémiai Kiadó |
| ISSN | 2062-509X (Print) |
| ISSN | 2062-8633 (Online) |
Monthly Content Usage
| Abstract Views | Full Text Views | PDF Downloads | |
|---|---|---|---|
| Oct 2022 | 4 | 0 | 0 |
| Nov 2022 | 2 | 0 | 0 |
| Dec 2022 | 4 | 0 | 0 |
| Jan 2023 | 9 | 0 | 0 |
| Feb 2023 | 9 | 0 | 0 |
| Mar 2023 | 4 | 0 | 0 |
| Apr 2023 | 0 | 0 | 0 |
Copyright Akadémiai Kiadó
AKJournals is the trademark of Akadémiai Kiadó's journal publishing business branch.
Copyright Akadémiai Kiadó AKJournals is the trademark of Akadémiai Kiadó's journal publishing business branch.
Powered by PubFactory