Improved detection of human antibodies to a Plasmodium antigen using a peptide modified with Aib residues.
A 17-mer sequence was selected as a model to study the influence of modifications of terminal ends both on the conformational of a peptide and on its antigenicity towards naturally developing antibodies. This sequence corresponded to a tandemly repeated motif, found in a long repetitive region, with high helical propensity, of a Plasmodium falciparum liver-stage antigen (LSA-1), immunogenic in man. Our model peptide was synthesized with ionizable or non-ionizable ends, or modified in both extremities by introduction of the helix-promoting residue alpha-aminoisobutyric acid (Aib). Helical contribution, absent in the 17 amino-acid sequence possessing ionizable ends, was detectable when non-ionizable ends were introduced, and dramatically increased in the Aib-modified analogue. The presence of ionizable ends totally abolished reactivity towards human sera, otherwise detectable with the peptide possessing non-ionizable ends. While modification by Aib residues was neither detrimental nor beneficial to antigenicity in solution, it clearly resulted in an improved sensitivity of the specific antibody detection when used as solid-phase antigen in ELISA.
Bossus
M
M
Service de chimie des biomolécules, URA CNRS 1309, Institut Pasteur de Lille, France.
BenMohamed
L
L
Londono
A
A
Barbier
B
B
Tartar
A
A
Druilhe
P
P
Gras-Masse
H
H
Mohammed
L
L
D000595:Amino Acid Sequence; D000621:Aminoisobutyric Acids / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000913:Antibodies, Protozoan / Q000737:chemistry*; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology*; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D010963:Plasmodium falciparum / Q000276:immunology*; D011487:Protein Conformation
D000621:Aminoisobutyric Acids; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D010455:Peptides; C067302:liver stage-specific antigen, Plasmodium; C100049:2-aminoisobutyric acid