title: Rational design and evaluation of a multiepitope chimeric fusion protein with the potential for leprosy diagnosis.
abstract: Despite the reduction in the number of leprosy cases registered worldwide as a result of the widespread use of multidrug therapy, the number of new cases detected each year remains stable in many countries. This indicates that Mycobacterium leprae, the causative agent of leprosy, is still being transmitted and that, without an earlier diagnosis, transmission will continue and infection will remain a health problem. The current means of diagnosis of leprosy is based on the appearance of clinical symptoms, which in many cases occur after significant and irreversible nerve damage has occurred. Our recent work identified several recombinant antigens that are specifically recognized by leprosy patients. The goal of the present study was to produce and validate the reactivity of a chimeric fusion protein that possesses the antibody binding properties of several of these proteins. The availability of such a chimeric fusion protein will simplify future test development and reduce production costs. We first identified the antibody binding regions within our top five antigen candidates by performing enzyme-linked immunosorbent assays with overlapping peptides representing the amino acid sequences of each protein. Having identified these regions, we generated a fusion construct of these components (protein advances diagnostic of leprosy [PADL]) and demonstrated that the PADL protein retains the antibody reactivity of the component antigens. PADL was able to complement a protein that we previously produced (the leprosy IDRI [Infectious Disease Research Institute] diagnostic 1 [LID-1] protein) to permit the improved diagnosis of multibacillary leprosy and that had a good ability to discriminate patients with multibacillary leprosy from control individuals. A serological diagnostic test consisting of these antigens could be applied within leprosy control programs to reduce transmission and to limit the appearance of leprosy-associated disabilities and stigmatizing deformities by directing treatment.
authors.affiliation: Protein Advances Inc., 1102 Columbia St., Seattle, WA 98104, USA. malcolm.duthie@proteinai.com
authors.forename: Malcolm S
authors.initials: MS
authors.lastname: Duthie
authors.forename: Marah N
authors.initials: MN
authors.lastname: Hay
authors.forename: Cecile Z
authors.initials: CZ
authors.lastname: Morales
authors.forename: Lauren
authors.initials: L
authors.lastname: Carter
authors.forename: Raodoh
authors.initials: R
authors.lastname: Mohamath
authors.forename: Lucia
authors.initials: L
authors.lastname: Ito
authors.forename: Luiza K M
authors.initials: LK
authors.lastname: Oyafuso
authors.forename: Marli I P
authors.initials: MI
authors.lastname: Manini
authors.forename: Marivic V
authors.initials: MV
authors.lastname: Balagon
authors.forename: Esterlina V
authors.initials: EV
authors.lastname: Tan
authors.forename: Paul R
authors.initials: PR
authors.lastname: Saunderson
authors.forename: Steven G
authors.initials: SG
authors.lastname: Reed
authors.forename: Darrick
authors.initials: D
authors.lastname: Carter
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology; D019411:Clinical Laboratory Techniques / Q000379:methods*; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007918:Leprosy / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D009166:Mycobacterium leprae / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D055815:Young Adult
chemical_list: D000942:Antigens, Bacterial; D000939:Epitopes; D011993:Recombinant Fusion Proteins