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AM [no-re active (Xenrrbindl

<1%

<1

CMT-GAPS 2 am ¡no -si 1 arre coaterl slides (Corningl

<1%

<1

Figure 13.2 Evaluation of anti-peptide affinity array production parameters. (a) Choice of substrate: data obtained by spotting fluorescently labelled albumin onto untreated substrates. The values reflect not only the protein binding capacity of the substrate but also the fluorescence quenching by the substrate. (b) An example of the best substrate: Nylon membrane, immobilized onto a glass slide. (c) Fluorescence quenching by other (non-traditional) substrates. (d) The effect of crosslinking on signal stability (following blocking and washing. (e) Untreated membrane: membranes were incubated in sealed beakers filled with undiluted formaldehyde (10 ml per 100-ml beaker volume) overnight

Figure 13.2 Evaluation of anti-peptide affinity array production parameters. (a) Choice of substrate: data obtained by spotting fluorescently labelled albumin onto untreated substrates. The values reflect not only the protein binding capacity of the substrate but also the fluorescence quenching by the substrate. (b) An example of the best substrate: Nylon membrane, immobilized onto a glass slide. (c) Fluorescence quenching by other (non-traditional) substrates. (d) The effect of crosslinking on signal stability (following blocking and washing. (e) Untreated membrane: membranes were incubated in sealed beakers filled with undiluted formaldehyde (10 ml per 100-ml beaker volume) overnight samples. Note that fewer antibodies could have been used to achieve a correct match.

The effect of sample collection and storage conditions on the sample stability has been discussed in the literature for many years, mostly in relation to clinical chemistry applications. Affinity peptidomics is the most generic affinity assay system to date, applicable for protein identification, quantification and expression profiling. In its concept it is the opposite of the 'sample preservation' strategies employed or attempted in traditional biomolecular diagnostics. It requires complete proteolytic digestion of the proteinaceous samples as the first step of any analysis. It is therefore suitable for the analysis of inconsistently stored or partially degraded samples, hence is suitable for a wide range of application, ranging from routine biomedical diagnostics to biometrics and forensics analyses. Protein microarrays and peptidomics are the two enabling technologies that allow fast and accurate protein profiling and would yield a new alternative methodology that is likely to supersede traditional protein

Table 13.3 Simulation of forensic material (samples were stored for 9 months under the conditions specified)

Batch no.a

Storage

To simulate

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