A generic multi-component approach was designed to perform simultaneous in situ polymerization and ligand immobilization to develop affinity porous polymer based chromatography resin in a facile mode. This strategy exploits the regioselective ring opening reaction between epoxy group of monomer and native functional group of ligand (i.e. amine) under aqueous condition (pH 9.7). As a proof-of-concept, reaction of iminodiacetic acid (IDA) with allyl glycidyl ether (AGE) in presence of other monomer (HEMA) and crosslinkers (DATD, PDA) for 4 h via thermal initiation process (temperature of 65 °C) was shown. Successful polymerization (both ex situ &amp; in situ) was confirmed by visual observation, surface morphology of the polymer by scanning electron microscope and ligand immobilization by FT-IR analysis. Chelation of the metal-ion i.e. copper (Cu (II)) with IDA in the monolith showed IgG adsorption capacity (27.8 mg/g monolith) over IDA-monolith without metal-ion. The affinity column has shown efficient capture of high abundant proteins such as IgG, transferrin and albumin from human plasma. © 2020 Elsevier B.V.

Kali Kishore Reddy Tetala

Centre for Bio-Separation and Technology

Vellore Campus

Khaparde A

Vellore Institute of Technology (VIT) is a private university located in&nbsp;Tamil Nadu, India. Founded in 1984, as Vellore Engineering College, the institution offers 20 undergraduate, 34 postgraduate, four integrated and four research programs. It has campuses in Vellore, Amravati, Bhopal and Chennai.

VIT is one of the top ranked private universities in India according to NIRF, THE and QS Rankings.&nbsp;Govt. of India has recognized&nbsp;VIT, Vellore as an&nbsp;Institution of Eminence. This has allowed VIT to take independent quality initiatives and move up in world ranking.

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VIT University

Journal of Pharmaceutical and Biomedical Analysis

In this study, we report the development of a new poly HEMA (HEMA-co-DEGDA-co-DATD) monolith capillary functionalized with “IDA-Cu (II) complex”. Of the two tested crosslinkers (methylene bisacrylamide (MBAAm) and diethylene glycol diacrylate (DEGDA)), presence of DEGDA has enhanced the monolith rigidity. Structural assembly of these monoliths are organized with highly interconnected large globule like structures and dominated by macropore region. Iminodiacetic acid (IDA) immobilization was performed using two chemical approaches (i. aldehyde − secondary amine reaction and ii. epoxy − sec. amine reaction). FT-IR analysis confirmed successful IDA immobilization in both cases. For the first time, a reaction of sec. amine ligand with aldehyde functional material was successfully reported. Overall, the Cu (II)-IDA monolith capillary showed good permeability (3.05 × 10 −13 m2), high IgG adsorption capacity and reusablilty even after 5 consecutive adsorption-desorption cycles. The amount of protein (IgG/HSA) adsorbed on Cu (II)-IDA monolith prepared via the two chemistries is almost similar. Using this affinity monolith capillary, we selectively depleted ∼95% of IgG from human plasma (dilution of 1:16). © 2017 Elsevier B.V.

Journal of Chromatography A

Development of a metal/chelate polyhydroxyethylmethacrylate monolith capillary for selective depletion of immunoglobulin G from human plasma for proteomics

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. © 2017 Elsevier B.V.

Journal of Chromatography B

A structure based plasma protein pre-fractionation using conjoint immobilized metal/chelate affinity (IMA) system

Immobilized metal-ion affinity (IMA) adsorption is a collective term that is used to include all kinds of adsorptions where the metal ion serves as the characteristic and most essential part of adsorption center. Of all the IMA techniques, immobilized metal-affinity chromatography (IMAC) has been gaining popularity as the choice of purification technique for proteins. IMAC represents a separation technique that is primarily useful for proteins with natural surface exposed-histidine residues and for recombinant proteins with engineered histidine tag. This review also gives insight into other nonchromatographic applications of IMA adsorption such as immobilized metal-ion affinity gel electrophoresis (IMAGE), immobilized metal-ion affinity capillary electrophoresis (IMACE), and immobilized metal-ion affinity partitioning (IMAP).

Journal	Data powered by TypesetJournal of Pharmaceutical and Biomedical Analysis
Publisher	Data powered by TypesetElsevier BV
ISSN	0731-7085
Open Access	No