Revisited BIA-MS combination: Entire “on-a-chip” processing leading to the proteins identification at low femtomole to sub-femtomole levels

Abstract

We present the results of a study in which biomolecular interaction analysis (BIA, Biacoretrade mark 2000) was combined with mass spectrometry (MS) using entire "on-a-chip" procedure. Most BIA-MS studies included an elution step of the analyte prior MS analysis. Here, we report a low-cost approach combining Biacore analysis with homemade chips and MS in situ identification onto the chips without elution step. First experiments have been made with rat serum albumin to determine the sensitivity and validation of the concept has been obtained with an antibody/antigen couple. Our "on-a-chip" procedure allowed complete analysis by MS/MS(2) of the biochip leading to protein identifications at low femtomole to sub-femtomole levels. Using this technique, identification of protein complexes were routinely obtained giving the opportunity to the "on-a-chip" processing to complete the BIA-MS approach in the discovery and analysis of protein complexes.

Qualitative and Quantitative Analysis of the Binding of GII.4 Norovirus Variants onto Human Blood Group Antigens

“…Since 2002, the new GII. 4 NoV strains have been characterized by conserved mutations in their capsids and the insertion of one amino acid at position 395 in the hypervariable P2 domain of the capsid proteins (12). This additional amino acid, which is mostly a threonine residue, has been conserved for all the GII.4 strains ever since the emergence of the Farmington variants.…”

“…4 NoVs now predominate over NoVs from other genogroups (6,46). However, little is known about the mechanisms that cause GII.4 variants to circulate for a couple of years before disappearing from the community.…”

“…The binding of purified VLPs to lacto-N-fucopentaose I (LNFP-I) and to A and B trisaccharides conjugated to BSA (Dextra Laboratories) ( Table 3) was analyzed by surface plasmon resonance at 25°C with a Biacore 2000 instrument (GE Healthcare). The homemade gold chips for SPR were designed and fabricated as described previously (4). The chemical functionalization was obtained by using a mixture of 11-mercapto-1-undecanol (11-MUOH) and 16-mercapto-1-hexadecanoic acid (16-MHA) (Sigma-Aldrich).…”

Membrane-associated Hsp72 from tumor-derived exosomes mediates STAT3-dependent immunosuppressive function of mouse and human myeloid-derived suppressor cells

“…Design and fabrication of homemade chips compatible with Surface Plasmon Resonance have been performed as previously published with the help of the MIMENTO technological platform (Besançon, France) (62). Biacore experiments were performed with the Biacore 2000 apparatus at 25°C with a flow rate of between 2 and 30 μl/min.…”

Surface plasmon resonance imaging in arrays coupled with mass spectrometry (SUPRA–MS): proof of concept of on-chip characterization of a potential breast cancer marker in human plasma

“…In this field, since 2000, pioneering studies have investigated MS analysis directly from biochips to detect whole proteins or peptides after in-situ enzymatic digestion of captured proteins [16-19, 21, 22]. It was, however, only recently that sensitive detection of analytes by MS and MS-MS in ideal solutions on arrays was efficiently achieved [23,24]. However, as far as we are aware, an analytical solution-based "on-a-chip" SPR-MS technique for characterization of protein biomarkers in biological samples has still to be developed and validated.…”