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  • Development of an improved fractionation of the human plasma proteome by a combination of abundant proteins depletion and multi-lectin affinity chromatography.

Development of an improved fractionation of the human plasma proteome by a combination of abundant proteins depletion and multi-lectin affinity chromatography.

Bioanalysis (2014-11-21)
Francisca O Gbormittah, Marina Hincapie, William S Hancock
ABSTRACT

Current analytical tools lack the required capacity to reduce the complexity of the plasma proteome and identify low-level proteins of clinical interest. Hence, the need to develop a fractionation approach to provide adequate throughput for a clinical study and minimize the loss and improve the detection of low abundance proteins. We present the development of an analytical platform that combines the depletion of 12 high abundance proteins and multi-lectin affinity chromatography (12P-M-LAC) fractionation. We validated the highly specific, stable and robust 12P-M-LAC platform using human plasma. An improved enrichment of low abundance proteins and glycoproteins with minimum sample loss was achieved demonstrating the suitability of this platform in future biomarker discovery studies.

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Sigma-Aldrich
Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
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Ethyl alcohol, Pure, 190 proof, for molecular biology
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Ethyl alcohol, Pure, 200 proof, ACS reagent, ≥99.5%
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Acetonitrile, HPLC Plus, ≥99.9%
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Acido formico, reagent grade, ≥95%
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HEPES, ≥99.5% (titration)
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HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
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Ethyl alcohol, Pure, 200 proof, meets USP testing specifications
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Acido formico, ACS reagent, ≥96%
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Acido formico, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥98%
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Acetonitrile, ACS reagent, ≥99.5%
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Acetonitrile, for HPLC, for UV, ≥99.9% (GC)
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Sodio cloruro, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
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Acetonitrile, suitable for HPLC, gradient grade, ≥99.9%
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Cloruro di calcio, BioUltra, for molecular biology, ~1 M in H2O
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Sodio cloruro, 0.9% in water, BioXtra, suitable for cell culture
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Sodio cloruro, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
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Sodio cloruro, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
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BIS-TRIS, ≥98.0% (titration)
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Acido formico, puriss., meets analytical specifications of DAC, FCC, 98.0-100%
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Cloruro di calcio, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%
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Divinylbenzene, technical grade, 80%
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Acido formico, ACS reagent, ≥88%
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HEPES, BioUltra, for molecular biology, ≥99.5% (T)
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Etanolo, ACS reagent, prima fine spirit, without additive, F15 o1
SAFC
Sodio cloruro, 5 M
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Etanolo, purum, absolute ethanol, denaturated with 4.8% isopropanol, A15 IPA1, ≥99.8% (based on denaturant-free substance)
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Soluzione tampone HEPES, 1 M in H2O
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Acetonitrile, suitable for HPLC-GC, ≥99.8% (GC)
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Sodium azide, BioUltra, ≥99.5% (T)