A structure based plasma protein pre-fractionation using conjoint immobilized metal/chelate affinity (IMA) system.
The potential of immobilized metal/chelate affinity (IMA) in a continuous fashion, referred as conjoint approach, to pre-fractionate plasma proteins (in their native state) prior to LC-MS analysis was investigated in this study. Four transition metal-ions (Co (II), Zn (II), Ni (II) and Cu (II)) were individually chelated with IDA (iminodiacetic acid) coated CIM (Convective Interaction Media) disks and placed in a single housing in the following sequential order: IDA-Co (II)→IDA-Zn (II)→IDA-Ni (II)→IDA-Cu (II). The rationale behind this order is to retain proteins based on their specific requirement for surface exposed histidine topography. This structural pre-fractionation hypothesis was successfully proven using four human plasma proteins (fibrinogen, IgG, transferrin, and albumin) with varying histidine topographies. This conjoint IMA pre-fractionation strategy not only fractionated proteins (from plasma) based on their native surface histidine topography, but also identified 157 proteins from human plasma. The advantage of our conjoint IMA is its ability to fractionate proteins in their native state and reduce plasma complexity in a single step by employing single buffer system.
Advanced Centre for Bioseparation Technology (CBST), VIT University, Vellore, Tamilnadu 632014, India.
Karan
K
Karkra
Advanced Centre for Bioseparation Technology (CBST), VIT University, Vellore, Tamilnadu 632014, India.
Kishore K R
KKR
Tetala
Advanced Centre for Bioseparation Technology (CBST), VIT University, Vellore, Tamilnadu 632014, India. Electronic address: indviji@yahoo.com.
M A
MA
Vijayalakshmi
D000327:Adsorption; D001798:Blood Proteins / Q000032:analysis / Q000302:isolation & purification*; D002614:Chelating Agents / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D006639:Histidine / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D007098:Imino Acids / Q000737:chemistry*; D008670:Metals / Q000737:chemistry*; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
D001798:Blood Proteins; D002614:Chelating Agents; D007098:Imino Acids; D008670:Metals; D006639:Histidine; C008109:iminodiacetic acid
CIM disk; Conjoint IMA; Plasma; Pre-fractionation; Proteins