ANNUAL REPORT - Department of Biotechnology

Atlanta and IISc., Bangalore-Stanford University,
USA, phase-I clinical trials of the rotaviral diarrhoea
vaccine strains (116E & I321) for the safety and
immunogencity studies have been completed in
India and USA using the vaccine produced in AGMK
cell lines in adults, older children and infants,
suggesting that the vaccine is safe. Vaccine take was
reported in 74% of recipients of 116E vaccine
candidate and 40% recipients of I321 vaccine. M/s.
Bharat Biotech International Ltd. (BBIL), Hyderabad
has produced prototype vero cell based vaccine
(116E), under cGMP conditions and initiated Double-
Blind Randomized Placebo Controlled Dose
Escalating Phase Ib/IIa study in December, 2006 to
evaluate the safety and immunogenicity of live
attenuated rotavirus vaccine 116E in healthy non
malnourished infants 8-20 weeks of age. As an initial
step, infants were screened for eligibility (healthy,
non-malnourished) for receiving the vaccine.
Subsequently rotaviral diarrhoea vaccine was
administered to eligible infants.
Malaria
Under the project implemented at ICGEB and
NMRC, New Delhi, expression of recombinant
PvRII (one of the P.vivax duffy binding protein)
encoding for 38 kD product has been studied. Pilot
scale (10L fermentation scale) methods to produce
correctly folded recombinant PvRII have been
developed with final yield of ~200 mg PvRII per L
fermentation culture and produced at pilot scale and
formulated with Montanide ISA 720, AS02A, QS21,
MF59 and alum has been used for immunogenicity
studies in mice. Formulations with Montanide ISA
720 and AS02A are highly immunogenic in mice and
elicit high titer binding inhibitory antibodies. Methods
for production of PvRII have been scaled up to 50L
fermentation scale and 100L refolding scale at the
Bharat Biotech International Ltd. under cGMP
conditions. Further stability studies have been
performed with two batches of recombinant PvRII
produced at 50L scale at two temperatures.
Lyophilization condition for recombinant PvRII has
been developed. Formulation of recombinant PvRII
with AS02A has been tested for stability where PvRII
was found to be stable in the emulsion. The scale up
studies and recombinant PvRII have been carried
out at BBIL, Hyderabad, including establishment of
downstream processes and various quality control
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parameters etc. Toxicology studies have been
completed and planned to start phase-I clinical trial
are being initiated in humans.
Hepatitis-C
The joint project implemented at Deccan College of
Medical Sciences & Allied Hospitals, Hyderabad and
University of Tennessee Health Sciences Center,
Memphis, USA, on molecular epidemiology of
genetic variation in the Hyper Variable Region-I
(HVR-I) sequence of Indian patients and response to
interferon, therapy, 300 HCV infected samples for
genotyping in north and south India (Hyderabad,
New Delhi, Lucknow) have been studied. Cloning
and sequencing of the complete genome of HCV
strain isolated from a single patient was carried out.
The complete sequence was deposited in GenBank
and can be retrieved using Accession No.:
AY051292. Further comparative studies have been
carried out with other known full-length sequences
isolated from various geographical regions and it was
observed that the Indian strains are closely related to
Indonesian strains and there is high heterogeneity on
gene sequences in envelope region.
Further twenty two of the 59 samples analyzed for
HVR region showed 100% homology to the
AY051292 strain, while only 4 showed 100%
similarity to the AY651061 strain. Under the present
study only 17 of the enrolled 25 patients, samples
showed response to the therapy. The study also
shows that genotype 3 samples had a response rate
of 77.7% when compared to genotype 1 samples that
had a response rate of 62.5%.
Tuberculosis
The collaborative study on “High throughput
PCR assays for diagnosing tuberculosis caused by
Mycobacterium tuberculosis and Mycobacterium
bovis using molecular beacons” at AIIMS, New Delhi,
CJILMD, Agra and Public Health Research Institute,
Newark, New Jersey supported with an objective to
develop reliable devR- and hupB-based PCR assays
in visual format using molecular beacons. The group
has collected a total of 77 sputum specimens from
LRSI of TB and Respiratory Diseases. The samples
were processed for culture and devR PCR by the