25 / 27
allied
academies
Page 62
Notes:
June 06-07, 2019 | London, UK
2
nd
International Conference on
Tissue Science and Molecular Biology,
Stem Cells & Separation Techniques
Joint Event
Biomedical Research (An International Journal of Medical Sciences) | ISSN: 0976-1683 Volume 30
Pseudobioaffinity ligands coupled to high throughput support matrix for
purification of proteins from preparative to analytical validation important aspect
in DSP of biopharma
M A Vijayalakshmi
Lab.interaction Moleculaire Tech, France
T
he term “Pseudobiospecific affinity chromatography” was
coined by Prof. MA Vijayalakshmi in 1989 (TIBTECH 1989),
which cover classes of ligands which have both structural and
functional recognition of proteins based on their aminoacid
sequence and three dimensional structure. Pseudobiospecific
ACsystems(e.g.aminoacids,metal-chelates andtriazinedyes)
are highly economic and robust, canbe fine-tuned to excellent
specificities and medium dissociation constants (10-7 - 10-5).
Ligand and Matrix are the two chromatography components
that guidemolecular interactions in any AC system. The ligand
governs thermodynamic aspect of the chromatographic
system which includes binding specificity, binding strength,
ligand concentration etc. The support matrix governs the
high-throughput-hydrodynamic aspect which includes the
porosity of the support, particle size, etc. “MONOLITHS”
are new stationary phase materials introduced in 1990’s as
“non-particulate homogeneous methacrylate material with
high pore interconnectivity and lack of interestial voids”
containing mega pores. They can be prepared in different
forms like disks (CIM®: Convective Interactive Media), radio
flow columns, capillaries and microfluidics. Due to the high
pore interconnectivity, the flow is convective which results
in efficient mass transfer of molecules and without any
diffusion limitation like in the agarose based system. Thus a
flow independent binding systemgives very high capacity and
binding, even at very high flow rates like 5 column volumes
per minute. The CIM systems are hydrophilic and versatile
such that any ligand can be coupled as is being done with
agarose matrices and with same chromatographic buffer
systems. Chromatographic runs are done seconds to minutes
not in hours and days. Apart from these, monoliths possess
other advantages like ease of preparation, low dead volumes,
chemical and mechanical stability and compatibility to get
hyphenated with conventional chromatography equipment’s.
e
:
indviji@yahoo.comBiomed Res, Volume 30
ISSN: 0976-1683