{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T01:44:16Z","timestamp":1781660656800,"version":"3.54.5"},"reference-count":49,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2017,11,22]],"date-time":"2017-11-22T00:00:00Z","timestamp":1511308800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Malaysian Ministry of Higher Education through the Higher Institution Centre of Excellence (HICoE) Grant","award":["311\/CIPPM\/4401005"],"award-info":[{"award-number":["311\/CIPPM\/4401005"]}]},{"name":"Universiti Sains Malaysia APEX DE Grant","award":["1002\/CIPPM\/910316"],"award-info":[{"award-number":["1002\/CIPPM\/910316"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Helminth parasite infections are significantly impacting global health, with more than two billion infections worldwide with a high morbidity rate. The complex life cycle of the nematodes has made host immune response studies against these parasites extremely difficult. In this study, we utilized two phage antibody libraries; the immune and na\u00efve library were used to identify single chain fragment variable (scFv) clones against a specific filarial antigen (BmR1). The V-gene analysis of isolated scFv clones will help shed light on preferential VDJ gene segment usage against the filarial BmR1 antigen in healthy and infected states. The immune library showed the usage of both lambda and kappa light chains. However, the na\u00efve library showed preferential use of the lambda family with different amino acid distributions. The binding characteristics of the scFv clones identified from this work were analyzed by immunoassay and immunoaffinity pull down of BmR1. The work highlights the antibody gene usage pattern of a na\u00efve and immune antibody library against the same antigen as well as the robust nature of the enriched antibodies for downstream applications.<\/jats:p>","DOI":"10.3390\/ijms18112376","type":"journal-article","created":{"date-parts":[[2017,11,22]],"date-time":"2017-11-22T10:47:38Z","timestamp":1511347658000},"page":"2376","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Antibody-Based Protective Immunity against Helminth Infections: Antibody Phage Display Derived Antibodies against BmR1 Antigen"],"prefix":"10.3390","volume":"18","author":[{"given":"Anizah","family":"Rahumatullah","sequence":"first","affiliation":[{"name":"Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Penang, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Izzati","family":"Abdul Karim","sequence":"additional","affiliation":[{"name":"Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Penang, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7583-0752","authenticated-orcid":false,"given":"Rahmah","family":"Noordin","sequence":"additional","affiliation":[{"name":"Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Penang, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0656-3045","authenticated-orcid":false,"given":"Theam","family":"Lim","sequence":"additional","affiliation":[{"name":"Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Penang, Malaysia"},{"name":"Analytical Biochemistry Research Centre, Universiti Sains Malaysia, Minden 11800, Penang, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,11,22]]},"reference":[{"key":"ref_1","first-page":"985680","article-title":"Lymphatic filariasis disseminating to the upper extremity","volume":"2014","author":"Maldjian","year":"2014","journal-title":"Case Rep. Radiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"363","DOI":"10.4269\/ajtmh.2000.62.363","article-title":"Application of a polymerase chain reaction-elisa to detect Wuchereria bancrofti in pools of wild-caught anopheles punctulatus in a filariasis control area in papua new guinea","volume":"62","author":"Bockarie","year":"2000","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1179\/136485910X12786389891407","article-title":"Induction of immunoglobulin G4 in human filariasis: An indicator of immunoregulation","volume":"104","author":"Adjobimey","year":"2010","journal-title":"Ann. Trop. Med. Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1038\/nri2199","article-title":"Protective immune mechanisms in helminth infection","volume":"7","author":"Anthony","year":"2007","journal-title":"Nat. Rev. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1016\/S1286-4579(01)01477-0","article-title":"How the immune system protects the host from infection","volume":"3","author":"Janeway","year":"2001","journal-title":"Microbes Infect."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/jid.2013.521","article-title":"Antibody phage display: Technique and applications","volume":"134","author":"Hammers","year":"2014","journal-title":"J. Investig. Dermatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/978-1-61779-974-7_5","article-title":"Construction of human naive antibody gene libraries","volume":"907","author":"Hust","year":"2012","journal-title":"Methods Mol. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.it.2010.11.005","article-title":"To B or not to B: B cells and the Th2-type immune response to helminths","volume":"32","author":"Harris","year":"2011","journal-title":"Trends Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5570","DOI":"10.4049\/jimmunol.0803254","article-title":"Molecular dissection of antibody responses against pneumococcal surface protein A: Evidence for diverse D-H-less heavy chain gene usage and avidity maturation","volume":"182","author":"Rohatgi","year":"2009","journal-title":"J. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1038\/nature07930","article-title":"Broad diversity of neutralizing antibodies isolated from memory B cells in HIV-infected individuals","volume":"458","author":"Scheid","year":"2009","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11651","DOI":"10.1128\/JVI.01762-08","article-title":"Profiling the specificity of neutralizing antibodies in a large panel of plasmas from patients chronically infected with human immunodeficiency virus type 1 subtypes B and C","volume":"82","author":"Binley","year":"2008","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Throsby, M., van den Brink, E., Jongeneelen, M., Poon, L.L.M., Alard, P., Cornelissen, L., Bakker, A., Cox, F., van Deventer, E., and Guan, Y. (2008). Heterosubtypic neutralizing monoclonal antibodies cross-protective against H5N1 and H1N1 recovered from human IgM(+) memory B cells. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0003942"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1016\/j.molimm.2015.07.040","article-title":"Delineation of BmSXP antibody V-gene usage from a lymphatic filariasis based immune scFV antibody library","volume":"67","author":"Rahumatullah","year":"2015","journal-title":"Mol. Immunol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.toxicon.2016.04.032","article-title":"Generation of a naive human single chain variable fragment (scFV) library for the identification of monoclonal scFV against Salmonella Typhi Hemolysin E antigen","volume":"117","author":"Lim","year":"2016","journal-title":"Toxicon"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"690","DOI":"10.4161\/mabs.25592","article-title":"Comparing CDRH3 diversity captured from secondary lymphoid organs for the generation of recombinant human antibodies","volume":"5","author":"Venet","year":"2013","journal-title":"mAbs"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3001","DOI":"10.1038\/nprot.2007.448","article-title":"Selection of human antibody fragments by phage display","volume":"2","author":"Lee","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"187","DOI":"10.4331\/wjbc.v3.i12.187","article-title":"Frontier of therapeutic antibody discovery: The challenges and how to face them","volume":"3","author":"Lu","year":"2012","journal-title":"World J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1046\/j.1423-0410.2000.7820072.x","article-title":"Introduction to antibody engineering and phage display","volume":"78","author":"Watkins","year":"2000","journal-title":"Vox Sang."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"980250","DOI":"10.1155\/2012\/980250","article-title":"scFV antibody: Principles and clinical application","volume":"2012","author":"Ahmad","year":"2012","journal-title":"Clin. Dev. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2381","DOI":"10.1007\/s10529-014-1635-x","article-title":"Principles and application of antibody libraries for infectious diseases","volume":"36","author":"Lim","year":"2014","journal-title":"Biotechnol. Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1007\/s10059-011-2268-8","article-title":"Antibodies against non-immunizing antigens derived from a large immune scFV library","volume":"31","author":"Moon","year":"2011","journal-title":"Mol. Cells"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.jim.2005.01.014","article-title":"Construction, evaluation and refinement of a large human antibody phage library based on the IgD and IgM variable gene repertoire","volume":"299","author":"Loset","year":"2005","journal-title":"J. Immunol. Methods"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.jbiotec.2010.09.945","article-title":"A human scFV antibody generation pipeline for proteome research","volume":"152","author":"Hust","year":"2011","journal-title":"J. Biotechnol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"8015","DOI":"10.4049\/jimmunol.0802375","article-title":"The heavy chain variable segment gene repertoire in chronic chagas\u2019 heart disease","volume":"183","author":"Grippo","year":"2009","journal-title":"J. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4313","DOI":"10.1128\/IAI.71.8.4313-4319.2003","article-title":"VH3 gene usage in neutralizing human antibodies specific for the Entamoeba histolytica Gal\/GalNAc lectin heavy subunit","volume":"71","author":"Tachibana","year":"2003","journal-title":"Infect. Immun."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1002\/prot.22293","article-title":"Multi-constraint computational design suggests that native sequences of germline antibody H3 loops are nearly optimal for conformational flexibility","volume":"75","author":"Babor","year":"2009","journal-title":"Proteins Struct. Funct. Bioinform."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1038\/342877a0","article-title":"Conformations of immunoglobulin hypervariable regions","volume":"342","author":"Chothia","year":"1989","journal-title":"Nature"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1038\/321522a0","article-title":"Replacing the complementarity-determining regions in a human-antibody with those from a mouse","volume":"321","author":"Jones","year":"1986","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1093\/protein\/gzt027","article-title":"The indistinguishability of epitopes from protein surface is explained by the distinct binding preferences of each of the six antigen-binding loops","volume":"26","author":"Kunik","year":"2013","journal-title":"Protein Eng. Des. Sel."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.jmb.2005.11.092","article-title":"Tyrosine plays a dominant functional role in the paratope of a synthetic antibody derived from a four amino acid code","volume":"357","author":"Fellouse","year":"2006","journal-title":"J. Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1016\/j.jmb.2003.10.007","article-title":"Expressed murine and human CDR-H3 intervals of equal length exhibit distinct repertoires that differ in their amino acid composition and predicted range of structures","volume":"334","author":"Zemlin","year":"2003","journal-title":"J. Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1186","DOI":"10.1039\/b927393j","article-title":"The functional capacity of the natural amino acids for molecular recognition","volume":"6","author":"Birtalan","year":"2010","journal-title":"Mol. Biosyst."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.sbi.2007.01.004","article-title":"The molecular architecture of protein-protein binding sites","volume":"17","author":"Reichmann","year":"2007","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/0014-5793(94)00648-2","article-title":"Amino-acid preferences at protein-binding sites","volume":"349","author":"Villar","year":"1994","journal-title":"FEBS Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1518","DOI":"10.1016\/j.jmb.2008.01.093","article-title":"The intrinsic contributions of tyrosine, serine, glycine and arginine to the affinity and specificity of antibodies","volume":"377","author":"Birtalan","year":"2008","journal-title":"J. Mol. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Barnes, M.R. (2007). Bioinformatics for Geneticists: A Bioinformatics Primer for the Analysis of Genetic Data, Wiley. [2nd ed.].","DOI":"10.1002\/9780470059180"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"31458","DOI":"10.1038\/srep31458","article-title":"Functional characterization of a monoclonal antibody epitope using a lambda phage display-deep sequencing platform","volume":"6","author":"Domina","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"10447","DOI":"10.1038\/srep10447","article-title":"Structural and functional characterization of a novel scFV anti-HSP60 of Strongyloides sp.","volume":"5","author":"Levenhagen","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_39","first-page":"302","article-title":"The structural basis of antibody-antigen recognition","volume":"4","author":"Kunik","year":"2013","journal-title":"Front. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"18067","DOI":"10.1074\/jbc.270.30.18067","article-title":"Crystal-structure of a cross-reaction complex between Fab F9.13.7 and guinea-fowl lysozyme","volume":"270","author":"Lescar","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"533","DOI":"10.4161\/mabs.27736","article-title":"3D modeling and characterization of the human CD115 monoclonal antibody H27K15 epitope and design of a chimeric CD115 target","volume":"6","author":"Grellier","year":"2014","journal-title":"MAbS"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Vangone, A., Abdel-Azeim, S., Caputo, I., Sblattero, D., Di Niro, R., Cavallo, L., and Oliva, R. (2014). Structural basis for the recognition in an idiotype-anti-idiotype antibody complex related to celiac disease. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0102839"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2285","DOI":"10.1093\/bioinformatics\/btt369","article-title":"Prediction of site-specific interactions in antibody-antigen complexes: The proabc method and server","volume":"29","author":"Olimpieri","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1080\/19420862.2016.1212149","article-title":"Phage display-derived human antibodies in clinical development and therapy","volume":"8","author":"Frenzel","year":"2016","journal-title":"MAbS"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1111\/j.1472-765X.2009.02694.x","article-title":"Effects of feeding and induction strategy on the production of BmR1 antigen in recombinant E-coli","volume":"49","author":"Norsyahida","year":"2009","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"D1006","DOI":"10.1093\/nar\/gkn838","article-title":"IMGT\u00ae, the international ImMunoGeneTics information system\u00ae","volume":"37","author":"Lefranc","year":"2009","journal-title":"Nucleic Acids Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"D671","DOI":"10.1093\/nar\/gki088","article-title":"VBASE2, an integrative V gene database","volume":"33","author":"Retter","year":"2005","journal-title":"Nucleic Acids Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"117","DOI":"10.2174\/157016461202150903114319","article-title":"Improved expression of single-chain fragment variable antibodies devoid of leader peptides in the cytoplasm","volume":"12","author":"Ismail","year":"2015","journal-title":"Curr. Proteom."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/B978-0-12-420119-4.00003-3","article-title":"Coupling antibody to cyanogen bromide-activated sepharose","volume":"Volume 541","author":"Kavran","year":"2014","journal-title":"Laboratory Methods in Enzymology: Protein, Pt C"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/11\/2376\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:50:43Z","timestamp":1760208643000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/11\/2376"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,11,22]]},"references-count":49,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2017,11]]}},"alternative-id":["ijms18112376"],"URL":"https:\/\/doi.org\/10.3390\/ijms18112376","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,11,22]]}}}