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Biomedical subjects

K S M S Raghavarao

Publications and source records attributed to K S M S Raghavarao.

6 recordsLinked to original sources

Method to obtain C-phycocyanin of high purity.

A new approach is made for the purification of C-phycocyanin (C-PC), which is simple and more efficient than existing methods. The proposed method involves two steps: aqueous two phase extraction and ion-exchange chromatography. Crude extract of C-phycocyanin, of purity 1.18, obtained from Spirulina platensis is subjected to aqueous two phase extraction. C-phycocyanin obtained from this process showed a purity of 5.22, which is higher than the reported value till date. In order to explore the possibility of further purification, C-phycocyanin is subjected to ion-exchange chromatography and found that the purity increased from 5.22 to 6.69. The fluorescence, intactness of structure and purity of C-phycocyanin are confirmed using spectrofluorometry, circular dichroism spectra and sodium dodecyl sulfate-polyacrylamide gel, respectively.

Chromatography, Ion Exchange↗

New aqueous two phase system comprising polyethylene glycol and xanthan.

A new aqueous two phase system comprising polyethylene glycol and xanthan is reported together with its phase diagram. The phase composition of the bottom phase did not vary (PEG 1.6-1.8% w/w; xanthan 0.24-0.28% w/w) while that of the top phase varied significantly (PEG 4-5% w/w, xanthan 0.05-1.37% w/w). Unlike conventional aqueous two phase systems, the viscosity of the top phase is also high and values are comparable to that of the bottom phase. When BSA was used as a model protein, it partitioned entirely into the bottom phase.

Animals↗

Drop formation in aqueous two-phase systems.

Extraction using aqueous two-phase systems (ATPSs) is a versatile technique for the downstream processing of various proteins/enzymes. The study of drop formation deals with the fundamental understanding of the behavior of liquid drops under the influence of various external body as well as surface forces. These studies provide a basis for designing of the extractions in column contactors in which liquid drops play a major role. Most of the drop formation studies reported so far is restricted to aqueous-organic systems. ATPSs, differ from aqueous-organic systems in their physical properties. In view of this, an attempt was made to develop a model for drop formation in ATPSs adopting the information available on aqueous-organic systems. In order to validate the model, experiments were performed by using polyethylene glycol (PEG)/salt systems of different phase compositions at various flow rates. At low flow rates the single stage model and at high flow rates the two stage model are able to predict the drop volume during its formation from tip of capillary. The experimental results were found to agree reasonably well with those predicted by the model.

Polyethylene Glycols↗

Microwave-field-assisted enhanced demixing of aqueous two-phase systems.

The slow rate of demixing is a major limitation in wide commercial exploitation of aqueous two-phase systems. In the present work, use of a microwave field has been explored for the first time to enhance phase demixing rates (decrease demixing times) of these systems. The microwave-field-assisted demixing process decreased the demixing time by about 2- to 4-fold in a polyethylene glycol/potassium phosphate system and by about 1.5- to 6.5-fold in a polyethylene glycol/maltodextrin system. The enhanced demixing rate can be explained by the dipole rotation, electrophoretic migration of free salts, multiple reflections at the interfaces, droplet-droplet collision, and reduced viscosity of the continuous phase that occur during the application of a microwave field.

Absorption↗

Acoustic field assisted demixing of aqueous two-phase systems.

Acoustic field assisted demixing was employed to decrease the demixing time in aqueous two-phase systems (polyethylene glycol-maltodextrin and polyethylene glycol-potassium phosphate). Application of acoustic field has decreased the demixing time in polyethylene glycol-maltodextrin by around twofold and up to about 3.2-fold in polyethylene glycol-potassium phosphate systems. Ultrasonication has induced mild circulation currents in the phase dispersion, which has enhanced the rate of droplet coalescence, eventually resulting in decreased demixing time. In the polyethylene glycol-maltodextrin system, phase demixing was found to depend greatly on which of the phases iscontinuous and viscosity of the continuous phase was observed to have a strong influence on the movement of the droplets and hence controlling the phase demixing rate. In case of the polyethylene glycol-potassium phosphate system, droplet coalescence was found to play a critical role in phase demixing. Addition of NaCl increased the demixing time and presence of Escherichia coli cells did not seem to have any influence on phase demixing.

Acoustics↗

Reverse micellar extraction for downstream processing of proteins/enzymes.

New developments in the area of downstream processing are, hopefully, to fulfill the promises of modern biotechnology. The traditional separation processes such as chromatography or electrophoresis can become prohibitively expensive unless the product is of high value. Hence, there is a need to develop efficient and cost-effective downstream processing methods. Reverse micellar extraction is one such potential and a promising liquid-liquid extraction technique, which has received immense attention for isolation and purification of proteins/enzymes in the recent times. This technique is easy to scale-up and offers continuous operation. This review, besides briefly considering important physico-chemical and biological aspects, highlights the engineering aspects including mass transfer, mathematical modeling, and technology development. It also discusses recent developments in reverse micellar extraction such as affinity based separations, enzymatic reactions in reverse micelles coupled with membrane processes, reverse micellar extraction in hollow fibers, etc. Special emphasis has been given to some recent applications of this technique.

Antigen-Antibody Reactions↗