A polypyrrole protein microarray for antibody–antigen interaction studies using a label-free detection process

Grosjean, Ludivine; Cherif, Boutheina; Mercey, Emilie; Roget, André; Lévy, Yves; Marche, Patrice N.; Villiers, Marie‐Bernadette; Livache, Thierry

doi:10.1016/j.ab.2005.09.033

Cited by 70 publications

(53 citation statements)

References 25 publications

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“…The technique used in this study to analyze the humoral responses of HDV-infected patients associates protein or peptide chips with SPRi detection, which allows the screening of large numbers of samples for numerous markers, including, for example, hepatitis B and/or hepatitis C serological analyses (20,23). However, the relevance of such an approach depends on the reproducibility of the measurements, and this point was first assessed for both the protein and peptide chips.…”

Section: Resultsmentioning

confidence: 99%

“…Pyrrole-protein conjugates were generated as described previously (20). Peptides were synthesized by Altergen (Bischheim, France) with a pyrrole-modified NH 2 terminus, as previously described (20). Pyrrole allows the grafting of probes on the gold surface of the prism array; it polymerizes under electricity power (21).…”

Section: Methodsmentioning

confidence: 99%

“…Proteins (10 M) or peptides (100 M) were grafted in triplicate on the gold surface of the biochip by electrochemical copolymerization, as described in Grosjean et al (20) and Cherif et al (22), respectively. The binding of specific Ab to probes (proteins or peptides) was monitored using surface plasmon res- onance imaging (SPRi).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Protein-Peptide Arrays for Detection of Specific Anti-Hepatitis D Virus (HDV) Genotype 1, 6, and 8 Antibodies among HDV-Infected Patients by Surface Plasmon Resonance Imaging

et al. 2015

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iLiver diseases linked to hepatitis B-hepatitis D virus co-or superinfections are more severe than those during hepatitis B virus (HBV) monoinfection. The diagnosis of hepatitis D virus (HDV) infection therefore remains crucial in monitoring patients but is often overlooked. To integrate HDV markers into high-throughput viral hepatitis diagnostics, we studied the binding of anti-HDV antibodies (Abs) using surface plasmon resonance imaging (SPRi). We focused on the ubiquitous HDV genotype 1 (HDV1) and the more uncommon African-HDV6 and HDV8 genotypes to define an array with recombinant proteins or peptides. Fulllength and truncated small hepatitis D antigen (S-HDAg) recombinant proteins of HDV genotype 1 (HDV1) and 11 HDV peptides of HDV1, 6, and 8, representing various portions of the delta antigen were grafted onto biochips, allowing SPRi measurements to be made. Sixteen to 17 serum samples from patients infected with different HDV genotypes were injected onto protein and peptide chips. In all, Abs against HDV proteins and/or peptides were detected in 16 out of 17 infected patients (94.12%), although the amplitude of the SPR signal varied. The amino-terminal part of the protein was poorly immunogenic, while epitope 65-80, exposed on the viral ribonucleoprotein, may be immunodominant, as 9 patient samples led to a specific SPR signal on peptide 65 type 1 (65#1), independently of the infecting genotype. In this pilot study, we confirmed that HDV infection screening based on the reactivity of patient Abs against carefully chosen HDV peptides and/or proteins can be included in a syndromebased viral hepatitis diagnostic assay. The preliminary results indicated that SPRi studying direct physical HDAg-anti-HDV Ab interactions was more convenient using linear peptide epitopes than full-length S-HDAg proteins, due to the regeneration process, and may represent an innovative approach for a hepatitis syndrome-viral etiology-exploring array.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Protein-Peptide Arrays for Detection of Specific Anti-Hepatitis D Virus (HDV) Genotype 1, 6, and 8 Antibodies among HDV-Infected Patients by Surface Plasmon Resonance Imaging

et al. 2015

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“…The generic method proposed here, based on the use of polypyrrole electrochemistry in picoliter volumes, was optimized for oligonucleotide probes but can be extended to a wide range of biological probes, provided some optimization in terms of concentration and buffer solutions are made to adapt existing protocols dedicated to more conventional electrochemical deposition. [22][23][24] The work presented here can therefore help to address some of the recent issues in surface patterning for generating biochips at reduced scales. It can be further improved, both in terms of resolution and diversity of deposited biomolecules, to meet specific requirements for screening purposes with enhanced sensitivity.…”

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confidence: 97%

Fabrication of Oligonucleotide Chips by Using Parallel Cantilever‐Based Electrochemical Deposition in Picoliter Volumes

et al. 2007

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Peptide Combinatorial Libraries

Liu

Lam

2008

Wiley Encyclopedia of Chemical Biology

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Peptide libraries, which consist of a few thousands to tens of millions of distinct peptides, can be generated combinatorially through biologic or synthetic approaches. The phage‐display peptide library method is simple and economical, and peptides with various natural protein folds can be generated by this approach; but this biologic method generally is limited to peptides that contain only L‐amino acids. Synthetic combinatorial libraries, however, can accommodate D‐amino acids, unnatural amino acids, and even organic moieties, which makes these approaches highly versatile. These synthetic methods include the spatially addressable parallel library method, the combinatorial library method that requires deconvolution, the one‐bead one‐compound (OBOC) combinatorial library method, the self‐assembled peptide nucleic acid (PNA) encoded chemical microarrays, and the synthetic library method that requires chromatography selection. Various methods to screen these combinatorial libraries have been developed. For example, phage‐display peptide libraries can be screened for the ability of phages to bind to unique immobilized target proteins or whole cells, peptide microarrays can be screened for their ability to interact with fluorescent proteins, combinatorial library methods that require deconvolution can be screened by standard solution‐phase assays, and OBOC libraries can be screened by on‐bead binding or functional assays or several solution‐phase cell‐based or biochemical assays. Combinatorial peptide library methods are enabling technologies that have proven to be very useful in basic research and drug discovery.

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A polypyrrole protein microarray for antibody–antigen interaction studies using a label-free detection process

Cited by 70 publications

References 25 publications

Protein-Peptide Arrays for Detection of Specific Anti-Hepatitis D Virus (HDV) Genotype 1, 6, and 8 Antibodies among HDV-Infected Patients by Surface Plasmon Resonance Imaging

Protein-Peptide Arrays for Detection of Specific Anti-Hepatitis D Virus (HDV) Genotype 1, 6, and 8 Antibodies among HDV-Infected Patients by Surface Plasmon Resonance Imaging

Fabrication of Oligonucleotide Chips by Using Parallel Cantilever‐Based Electrochemical Deposition in Picoliter Volumes

Peptide Combinatorial Libraries

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