In situ stimulation of a T helper cell hybridoma with a cellulose-bound peptide antigen

Ötvös, László; Pease, Anne Marie; Bokonyi, Krisztina; Giles-Davis, Wynetta; Rogers, Mark E.; Hintz, Paul A.; Hoffmann, Ralf; Ertl, Hildegund C. J.

doi:10.1016/s0022-1759(99)00194-5

Cited by 22 publications

(11 citation statements)

References 24 publications

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“…Most standard peptide synthesis equipment can synthesize only up to a few hundred single peptides simultaneously, although lately up to 8000 peptides have been synthesized in parallel on a cellulose membrane (23–25) using the SPOT TM technique. In addition to performing assays directly on the membrane (26), such peptides can be released and transferred onto glass slides using additional robotics and printing techniques (25). As alternatives to synthetic peptides, phages (27), bacteria (28), and yeast cells (29) have been used to express libraries of fragmented antigens (27) or of combinatorial peptides (30).…”

mentioning

confidence: 99%

High-resolution Mapping of Linear Antibody Epitopes Using Ultrahigh-density Peptide Microarrays

Buus

Rockberg

Forsström

et al. 2012

Molecular & Cellular Proteomics

183

174

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Antibodies empower numerous important scientific, clinical, diagnostic, and industrial applications. Ideally, the epitope(s) targeted by an antibody should be identified and characterized, thereby establishing antibody reactivity, highlighting possible cross-reactivities, and perhaps even warning against unwanted (e.g. autoimmune) reactivities. Antibodies target proteins as either conformational or linear epitopes. The latter are typically probed with peptides, but the cost of peptide screening programs tends to prohibit comprehensive specificity analysis. To perform high-throughput, high-resolution mapping of linear antibody epitopes, we have used ultrahigh-density peptide microarrays generating several hundred thousand different peptides per array. Using exhaustive length and substitution analysis, we have successfully examined the specificity of a panel of polyclonal antibodies raised against linear epitopes of the human proteome and obtained very detailed descriptions of the involved specificities. The epitopes identified ranged from 4 to 12 amino acids in size. In general, the antibodies were of exquisite specificity, frequently disallowing even single conservative substitutions. In several cases, multiple distinct epitopes could be identified for the same target protein, suggesting an efficient approach to the generation of paired antibodies. Two alternative epitope mapping approaches identified similar, although not necessarily identical, epitopes. These results show that ultrahigh-density peptide microarrays can be used for linear epitope mapping. With an upper theoretical limit of 2,000,000 individual peptides per array, these peptide microarrays may even be used for a systematic validation of antibodies at the proteomic level.

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mentioning

confidence: 99%

High-resolution Mapping of Linear Antibody Epitopes Using Ultrahigh-density Peptide Microarrays

Buus

Rockberg

Forsström

et al. 2012

Molecular & Cellular Proteomics

183

174

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“…Synthetic 15-mer peptides (v1 to v11) covering the HveC(143t) domain were generated so that they overlapped by 5 amino acids (v1, Q31 to G44 with an additional D at the N terminus; v2, Y40 to A54; v3, H50 to Q64; v4, V60 to S74; v5, S70 to S84; v6, I80 to R94; v7, L90 to F104; v8, L100 to L114; v9, R110 to C124; v10, G120 to R134; v11, P130 to M143). Peptides were synthesized on a polyethylene glycol-modified cellulose membrane as described previously (29). Membrane strips were probed overnight at 4°C with anti-HveC Ig (5 g/ml).…”

mentioning

confidence: 99%

Localization of a Binding Site for Herpes Simplex Virus Glycoprotein D on Herpesvirus Entry Mediator C by Using Antireceptor Monoclonal Antibodies

Krummenacher

Baribaud

Leon

et al. 2000

J Virol

110

138

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The human herpesvirus entry mediator C (HveC), also known as the poliovirus receptor-related protein 1 (PRR1) and as nectin-1, allows the entry of herpes simplex virus type 1 (HSV-1) and HSV-2 into mammalian cells. The interaction of virus envelope glycoprotein D (gD) with such a receptor is an essential step in the process leading to membrane fusion. HveC is a member of the immunoglobulin (Ig) superfamily and contains three Ig-like domains in its extracellular portion. The gD binding site is located within the first Ig-like domain (V domain) of HveC. We generated a panel of monoclonal antibodies (MAbs) against the ectodomain of HveC. Eleven of these, which detect linear or conformational epitopes within the V domain, were used to map a gD binding site. Among the 11 envelope glycoproteins of herpes simplex virus (HSV), glycoprotein D (gD) plays an essential role during viral entry into mammalian cells (14). gD binds specifically to one of several cell surface receptors during the pH-independent process that leads to fusion of the HSV envelope with the cell plasma membrane (13). Other essential glycoproteins such as gB and the gH-gL heterodimer also participate in the fusion event in ways that remain to be elucidated (9,35,38).Several HSV gD receptors have been identified. Herpesvirus entry mediator A (HveA; also known as HVEM and TNFRSF14) is a member of the tumor necrosis factor receptor family which binds gD and allows the entry of most HSV-1 and HSV-2 strains (25, 41). HveB (nectin-2) and HveC (nectin-1) are members of the immunoglobulin (Ig) superfamily that are closely related to the poliovirus receptor (PVR; also known as CD155) and to the newly discovered nectin-3 (8, 21, 22, 33). Whereas the activity of HveB is limited to certain HSV-2 strains and some laboratory strains of HSV-1 (rid1 and ANG) and pseudorabies virus (PRV) (20, 39), HveC allows the entry of all the HSV-1 and HSV-2 strains tested as well as PRV and bovine herpesvirus 1 (10). Poliovirus receptor does not function as an HSV receptor but can be used by PRV and bovine herpesvirus 1 (10). A specific type of heparan sulfate modified by D-glucosaminyl-3-O-sulfotransferase 3 can substitute for HveA or HveC and binds to gD to allow the entry of HSV-1 KOS into cells (34).HveB and HveC appear to be involved in cell-cell interaction and were named nectin-2 and nectin-1, respectively, according to their newly discovered function (1,19,37). In this paper, we will refer to them according to their viral usage (i.e., HveB and HveC).Recently, mutations in the HveC gene (named PVRL1 in that study) were linked to a form of cleft lip/palate-ectodermal dysplasia in humans (36).Although they have different structures, HveA and HveC bound to HSV-1 gD with similar affinity (17, 42). Using antibody competition and mutagenesis, the binding sites for HveC and HveA were mapped to common and distinct regions of gD (16,28,40). Reciprocally, the gD binding site on HveC has been localized to the first and most distal of the three Ig-like domains (or V domain) of its extr...

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“…Considering other peptide array studies, Otvos et al (18) have only reported the utilization of punched‐out peptide array for in situ stimulation of T‐helper cells without releasing peptides from the support. Their work had shown that peptides in SPOT peptide library are able to stimulate T‐cell by membrane‐bound form, and confirmed no release of peptides during the assay indicating that solid‐phase assay proceeds as true solid phase assay.…”

Section: Discussionmentioning

confidence: 99%

Pentamer peptide from Fas antigen ligand inhibits tumor‐growth with solid‐bound form found by peptide array

Kato

Kaga

Kunimatsu

et al. 2005

The Journal of Peptide Research

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Despite their low molecular weight and simple structure, peptides regulate vital reactions of various cells. With higher standards of safety and purity required in clinical researches, chemically synthesized peptides are considered as an ideal alternative material for animal‐derived materials in the regulation of cellular events. However, effective high‐throughput assay for studying peptide‐cell interactions has not been established to design peptide with objective function. Here, we report the effectiveness of the utilization of peptide array combined with cell assay for the design of cell‐interactive peptides. As a model case, peptide that regulates tumor cell viability with support‐bound form was explored. By culturing cells on peptide array, we found a novel 5‐mer sequence CNNLP (Cys‐Asp‐Asp‐Leu‐Pro) that strongly inhibits viability of tumor cells (leukemia and adenocarcinoma) from human Fas antigen ligand. We here indicate the advantageous features of peptide array, which are ‘feasibility in examining combinatorial amino acid substitutions’ and ‘indicative data consist of effective and ineffective substitutions from an assay’, contributes greatly for studying peptide‐cell interaction. These features differentiate peptide array from other peptide library screening strategy. As a result, we succeeded in obtaining 29 novel tumor‐growth inhibitory peptides with it solid‐bound form, and structural rules that suggest ideal peptide design for acquiring objective peptide function.

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In situ stimulation of a T helper cell hybridoma with a cellulose-bound peptide antigen

Cited by 22 publications

References 24 publications

High-resolution Mapping of Linear Antibody Epitopes Using Ultrahigh-density Peptide Microarrays

High-resolution Mapping of Linear Antibody Epitopes Using Ultrahigh-density Peptide Microarrays

Localization of a Binding Site for Herpes Simplex Virus Glycoprotein D on Herpesvirus Entry Mediator C by Using Antireceptor Monoclonal Antibodies

Pentamer peptide from Fas antigen ligand inhibits tumor‐growth with solid‐bound form found by peptide array

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