Biochemistry & Pharmacology: Open Access
Open Access
ISSN: 2167-0501
+44-20-4587-4809
ISSN: 2167-0501
+44-20-4587-4809
Mladen Krajacic
University of Zagreb, Croatia
Scientific Tracks Abstracts: Biochem Pharmacol (Los Angel)
The particular structure of monolithic chromatographic supports has been proved highly advantageous in analytic separation
and purification of large biomolecules. Characterized by very high porosity, high binding capacity, and high flow rate based
on convective mass transport, monoliths are particularly applicable in virus research. Monolith chromatography has hardly any
limitation to be applied in processing of virus particles, huge macromolecular complexes, and viral genomes, especially when
being distinct from nucleic acid forms present in the host cell. The majority of trials published so far, have exploited ion exchange
carriers, although other chemistries are also applicable, like hydrophobic interaction when concentrating viruses from marine
environment. According to papers published over the past decade, viruses of different sizes, structures and morphologies,
even virus-like particles, have successfully been purified from tissue homogenates or cell lysates. Moreover, following virtual
separation by overlapping chromatograms obtained from separate experiments, a real separation of three virus species, and
a distant strain of one of them (four viral fractions in total) was accomplished from laboratory prepared mixture. One of
the most recent achievements is a proof of principle that virus chromatographic feature could easily be modified without
abolishing its structural stability, or its biological activity. As adenoviruses have been used in almost a quarter of human gene
therapy trials, the experiment was conducted with an adenoviral vector. The deletion of just two negatively charged amino
acids from the main structural protein was efficient in shortening chromatographic retention of the recombinant adenovirus.
In this way, it would be possible to shift virus particles away from particular interfering substances present in the crude lysate.
The concept might be followed to facilitate chromatography-approach purification by engineering modifications contributing
to virus separability, besides those contributing to its therapeutic functionality.
Recent Publications
1. Krajacic M, Ravnikar M, Strancar A and Guti�?©rrrez-Aguirre I (2017) Application of monolithic chromatographic support
in virus research. Electrophoresis 38:2827-2836.
2. Ruscic J, Ambriovic-Ristov A, Majhen D, Kolundzija S, Barut M, Benihoud K and Krajacic M (2016) Manipulating
adenoviral vector ion-exchange chromatography: Hexon versus fiber. Journal of Separation Science 39:4299-4304.
3. Ruscic J, Guti�?©rrrez-Aguirre I, Tusek Znidaric M, Kolundzija S, Slana A, Barut M, Ravnikar M and Krajacic M (2015)
A new application of monolithic supports: The separation of viruses from one another. Journal of Chromatography A
1388:69-78.
4. Ruscic J, Guti�?©rrrez-Aguirre I, Urbas L, Kramberger P, Mehle N, Skoric D, Barut M, Ravnikar M and Krajacic M (2013) A
novel application of methacrylate based short monolithic columns: Concentrating Potato spindle tuber viroid from water
samples. Journal of Chromatography A 1274:129-136.
5. Krajacic M, Ivancic-Jelecki J, Forcic D, Vrdoljak A and Skoric D (2007) Purification of plant viral and satellite doublestranded
RNAs on DEAE monoliths. Journal of Chromatography A 1144:111-119.
Mladen Krajacic is a Full Professor at University of Zagreb where he completed his PhD in Virology. As a Head of the Laboratory and Principal Investigator in several research projects, he has been leading the molecular biology investigations on viruses, sub-viral agents (viroids, satellite RNAs) and non-cultivable bacteria. During the last decade, he has dedicated to chromatography on monolithic supports and its appliance in separation of viral and sub-viral ribonucleic acids, as well as virus particles. His results have been published in prominent scientific journals in which he has regularly been employed as a Reviewer. He was supervising a number of young research fellows who have received positions from well recognized institutions across the world.
E-mail: mladen.krajacic@biol.pmf.hr