4th International Conference and Exhibition on Biologics and Biosimilars October 26-28, 2015 Baltimore, USA

Awantika Singh is pursuing her PhD since July, 2011 from CSIR- Central Drug Research Institute, Lucknow and registered in Academy of Scientific and Innovative Research, New Delhi. She is senior research fellow (University Grant Commission, New Delhi) and her topic of research is qualitative and quantitative analysis of Indian medicinal plants and their herbal product using liquid chromatography coupled with tandem mass spectrometry. She has published 11 research articles in the reputed journals.

Berberis species are well known and used extensively as medicinal plants in traditional medicine. They have many medicinal values attributable to the presence of alkaloids having different pharmacological activities. In this study , a method was developed and validated as per international conference on harmonization guidelines using ultra high performance liquid chromatography with hybrid triple quadrupole-linear ion trap mass spectrometry operated in the multiple reaction monitoring mode for nine bioactive compounds, including protoberberine alkaloids, aporphine alkaloids and chlorogenic acid. This method was applied in different plant parts of eight Berberis species to determine variations in content of nine bioactive compounds. The separation was achieved on an ACQUITY UPLC CSH™ C18 column using a gradient mobile phase at flow rate 0.3 mL/min. Calibration curves for all the nine analytes provided optimum linear detector response (with R20.9989) over the concentration range of 0.5–1000 ng/mL. The precision and accuracy of the samples were within RSDs ≤2.4 and ≤2.3%, respectively. The results indicated significant variation in the total contents of the nine compounds in Berberis species. Results indicate that all the nine bioactive compounds are the most abundant in B. petiolaris stem. Chlorogenic acid, magnoflorine and berberine are found abundant in leaf, stem and root part, respectively of all the Berberis species. Results also indicated that the roots of B. lycium, B. koehneana and B. chitria may be used for swapping of B. aristata. This information was lacking in the literature and it could be helpful for better swapping of Berberis species.

Andreja Livk has over 17 years of experience in protein and peptide chemistry, including isolation and characterisation of peptides by RP-HPLC. Her skills include the synthesis of a wide range of peptides including complex bridged peptides. She has also extensively characterised low molecular mass peptides using a wide range of mass spectrometry systems, as well as de novo peptide sequencing for interpretation of MS/MS spectra.

A comprehensive physicochemical characterisation of novel biosimilar products is a prerequisite for their application and manufacturing. A side-by-side analysis, relative to reference products, can provide comparative data for biosimilars and demonstrate their suitability in relation to regulatory requirements. The aim of this study is to determine physicochemical properties of human insulin and its two analogues, aspart and glargine. In the insulin analogues, different amino acid residues are added or substituted at certain positions along the A and B chains to give them special acting characteristics in the human body. Mass spectrometry based peptide mapping, N/C terminal sequencing and intact mass analysis were conducted to determine differences in primary structures of the insulin analogues. Disulphide bond pairing, which is critical for protein’s structure and its function, was performed by online LC/ESI-MS/MS analysis. By mapping the position of each disulphide bond through sequential reduction and alkylation steps, mass spectrometry analysis eliminated downstream functional characterisation issues by confirming that the molecule is folded correctly.

Mitsuru Takahashi has completed his Ph.D at the age of 26 years from Tokyo Institute of Technology. He is working for Terumo Corporation for 5 years. He is the assistant manager of Technology Development of Global Pharmaceutical Solutions group.

The biotechnology sector continues to move towards the development of highly concentrated solutions as a way of providing a better response within the human body. There are potential issues of low injectability for high viscosity drugs. Tapered needle technology will be one of the solutions for improving injectability. mAb drug product (Omalizmab) was reconstituted with various volumes of WFI to attain appropriate viscosities of 269.6 cPs. Three needles (29G-24G tapered needle, 29G straight needle and 24G straight needle) were compared using four different viscosities mentioned above. The syringes used in the study were Terumo PLAJEXTM 1mL COP with i-coatingTM stopper. The syringes were filled with 1mL. The reduction rate of 29G-24G tapered needle at the speed of 6mL/min of the injection force was 46.2% when compared with the same gauge straight needle of 29G (The result was 65.01±1.43 N). Tapered needle technology reduced the injection force by approximately 46% when compared with same gauge straight needle.

Vikas Bajpai is pursuing his PhD from CSIR- Central Drug Research Institute, Lucknow and registered in Academy of Scientific and Innovative Research, New Delhi. He is senior research fellow (Council of Scientific and Industrial Research, New Delhi). He has more than six years’ experience in working and handling with many hyphenated mass spectrometric techniques. Currently he is engaged with the development of mass spectral fingerprints of natural products to check the different level of variations, adulteration from intact plant part using DART TOF MS technique.

Thalictrum (Ranunculaceae) is very widely distributed genus of medicinal plants in India and has been reported to have many therapeutic properties like diuretic, stomachic, antiseptic, aperient etc. Approximately 290 alkaloids were reported from the 80 species of the genus Thalictrum. Alkaloids from Thalictrum are reported to exhibit various pharmacological activities like antiamebic, antimicrobial, antitumour and antiviral. Protoberberine and aporphine alkaloids are reported the major components in this plant In present study, we present simultaneous quantification of some alkaloids in Thalictrum reniforme using ultra performance liquid chromatography coupled to hybrid triple quadrupole/linear ion trap mass spectrometry (UPLC-QqQLIT-MS) operated in the multiple reactions monitoring acquisition. The separation was achieved on an ACQUITY UPLC CSHâ„¢ C18 column using a gradient mobile phase at flow rate of 0.3 mL/min. Stock solutions of standards were prepared in methanol within the ranges from 0.5 to 1500 ng/mL. The calibration curves were constructed by plotting the peak areas versus the concentrations of each analyte. Five PBAs were quantified in which Berberine, Jatrorrhizine and Palmatine were detected in significant amount.

Siqin Liu is a PhD student in the Natural Drug Discovery group, School of Pharmacy, Queen’s University Belfast, UK. His current issue is peptides from skin secretion of amphibians, which is about finding and analyzing antimicrobial peptides, neuropeptides and enzyme relevant peptides in genetic and molecule levels. He also finished the bachelor degree in Beijing University of Chinese Medicine, China, which was majored about natural resource of Chinese herb medicine. He is handling technologies of real-time PCR, chromatography, cell line culture, next generation sequencing, etc.,

Skin secretions of amphibians, including frogs, toads and salamanders, are considered as abundant resources of bioactive peptides, which possess variety bio-functions, such as antibacterial, anti-inflammation, neuroreceptor antagonism and tumor inhibition. In especial, the feature of broad spectrum antimicrobial reveals bio-therapeutic potential. Act by creating transmembrane pores through the lipid bilayer, the alpha-helical short peptides split the microorganism based onthe concentration accumulation. This mechanism avoids the drug resistance effects which are led by receptor recognition. Consequently, the natural antimicrobial peptides emerge potential as alternative and supplement candidates in antibiotics agents. Brevinin-2 is a group of antimicrobial peptides whose members are typical helix and contain approximately 30 amino acid residues and a disulfide bond on the carboxyl terminus. In this study, structural modifications were created by replace residues at specific positions into lysine containing cationic side-chains. The modified peptide analogues revealed increase, in different levels, ofantimicrobial activity and tumor suppresser ability, along with unchanged or decreased haemolytic effect. These strategies redesigned the peptides structures, for instance, clustered the cationic residues from amphipathic helical and enhanced the hydrophobic property. The outcomes contributed the principles of optimizing transmembrane ability and reducing cytotoxic property of alpha-helical peptides, and made preparation of the further researches about therapeutic.

Tedla Mindaye, MSc, MLS (ASCPi)CM, has a degree in Medical Laboratory Science from the Addis Ababa University of Ethiopia (2006), specialization degrees in Tropical and Infectious Disease (2011) and currently is a PhD student. He was the Director of the Ethiopian Public Health laboratory Association (2012–2014). He has also serving as international ambassador for American Society for Clinical Pathology for international certification program since 2010. He has also participated as a speaker at national and international conferences and seminars and has been published a number of scientific articles in international journals.

Despite the fact that Ethiopia is accelerating its antiretroviral treatment (ART) program, little is known about the patient satisfaction on ART monitoring laboratory services in health facilities. The aim of the study is to assess patient satisfaction of ART monitoring laboratory services in public hospitals of Addis Ababa, Ethiopia. Hospital based, descriptive cross sectional study was conducted from October to November 2014 among clients attending in nine public hospitals in Addis Ababa Ethiopia. Patients’ satisfaction towards ART monitoring laboratory services was assessed using exit interview using structured questionnaire. Data were coded and entered using EPI info 2002 and analyzed using SPSS version 15 software. A total of 406 clients were involved in the study .Of these 255(62.8%) were females. The overall satisfaction rate for ART monitoring laboratory services was (85.5%). Patients were satisfied with measures taken by health care providers to keep confidentiality and ability of the person drawing blood to answer question (98.3% and 96.3% respectively). Moreover, the finding this study revealed that there was statistical significant associations between the overall patients’ satisfaction with waiting time to get blood drawing service, availability of ordered laboratory tests and waiting time to get laboratory result with (p<0.05). Patients receiving blood drawing service less than 30 minute were 7.59 times ( 95% CI AOR: 3.92-14.70) to be more satisfied with the ART monitoring laboratory services compared to those who underwent for more than 30 minutes.

Salimeh is a PhD student working under the supervision of Assoc. prof. Dr. Munir Abdul Murad. She received her Master’s degree in Biotechnology from the University Technology Malaysia. Her main research interests are protein engineering, enzymolgy, protein modeling and mamalian cell culture. Salimeh also cooperates with the Malaysion Genomic institute in the Framework for structural genomic project.

Glaciozyma antarctica PI12 is a psychrophilic yeast that was isolated from the surface of Antarctica sea ice. Psychrophiles thriving permanently at near-zero temperatures synthesize cold-active enzymes to sustain their survivals. Most psychrophilic enzymes are characterised by high catalytic efficiencies at low temperatures compared with their mesophilic counterparts. β-glucanases are carbohydrate acting enzymes which break down glycosidic bonds within β-glucan, which is usually present in plants and algae cell wall and play an important role in biodegradation of polysaccharide in nature. This study is the first to describe the molecular cloning, expression, purification and biochemical characterisation of novel cold active β-glucanase from psychrophilic yeast, G. antaractica PI12. cDNA sequence of endo-1,3(4)-glucanase (GaEGL) with an open reading frame of 1182 bp encoded for 394 amino acids has been cloned and expressed in E. coli expression system. Sequence analysis through bioinformatics studies revealed few homology identities of this protein with other endo-glucanases which belongs to glycosyl hydrolase family 16. The recombinant GaEgl with molecular mass of 44 kDa was purified with nickel affinity chromatography and subjected to enzymatic characterisation. Generally, this enzyme was active at low temperature and broad range of pHs, and able to hydrolyse several substrates such as laminarin, lichenan and yeast beta glucan. Analysis of the reaction products released using purified proteins on respective substrates via HPLC revealed that the main hydrolysis products were disaccharides and oligosaccharides. Overall, the findings of this study provide new insights on β-1,3(4)-glucanase from psychrophilic organism.

Ahmed Hassen graduated in pharmacy and joined the staff of pharmaceutics and industrial pharmacy department in College of Pharmacy, Cairo University, Egypt. He did his master degree in controlled release delivery. Currently he is a Ph.D scholar researching on transdermal drug delivery. His research interest focuses in drug delivery and bioequivalence studies.

Objective: The aim of this study was to assess the bioequivalence of two commercial 10-mg tablet formulations of rizatriptan, a selective 5-hydroxytryptamine1B/1D (5-HT1B/1D) receptor agonist indicated for the acute treatment of migraine attacks using a newly developed and validated LC-MS/MS assay. Methods: A simple, sensitive and specific LC-ESI/MS method was developed and validated for the determination of rizatriptan in human plasma in positive mode using the transitions m/z 270.6 --> 201.6 for rizatriptan and m/z 384.07 --> 100.1 for the internal standard. Rizatriptan and the internal standard were isolated from plasma samples by liquid-liquid extraction. The chromatographic separation was accomplished on a Luna C18 (phenomenex) (50 x 4.6) mm, 5μm with mobile phase consisting of acetonitrile: ammonium acetate (70:30 v/v) and 0.1% formic acid. A bioequivalence study was carried out in 24 healthy adult male volunteers using a single-dose, randomized, 2-way crossover design under fasting conditions. Both the test product and the reference product were administered after an overnight fast on two treatment days, separated by a 2-week washout period. After dosing, serial blood samples were collected for 12 hours. Statistical analysis of the pharmacokinetic parameters Cmax, AUC0-12, AUC0-∞ was conducted to determine bioequivalence. Results: Intraday and Interday assay precision was acceptable. The lower limit of quantification was 0.1 ng/mL. Accuracy was observed over a linear range of 0.1-100 ng/mL. The differences between the two products did not reach statistical significance with 90% CIs of 91.3-112.6, 101.6-111.2 and 102.0-111.6 for Cmax, AUC0-12 and AUC0-∞ respectively. The test/reference ratio of these parameters was within the acceptance range of the FDA criterion for bioequivalence. Both formulations were apparently well absorbed from the gastrointestinal tract (ie, no specific gastrointestinal tract-related adverse events were reported). Conclusions: The validated method offered increased sensitivity and wide linear concentration range. This method was successfully adopted for the evaluation of bioequivalence of two rizatriptan products after single dose administration to 24 volunteers.

Sunil Kumar is pursuing his PhD since Jan, 2013 from CSIR- Central Drug Research Institute, Lucknow and registered in Jawaharlal Nehru University, New Delhi. He is senior research fellow (CSIR, New Delhi) and his topic of research is qualitative and quantitative analysis of Indian medicinal plants and their herbal product using liquid chromatography coupled with mass spectrometry. He has published 4 research articles in the reputed journals.

Medicinal plants of the genus Rauwolfia (Apocynaceae) are extensively used as folk medicines worldwide. Its antihypertensive activity is well known due to the presence of monoterpene indole alkaloids (MIAs). The therapeutic potential of the herbal medicines are affected due to variation of bioactive phytoconstituents. Therefore, a rapid and validated method was developed for fingerprinting of roots and leaves of six Rauwolfia species by direct analysis in real time mass spectrometry (DART-MS). Seventeen bioactive MIAs were tentatively identified on the basis of their exact mass measurement from the intact plant parts. Further, principal component analysis (PCA) was used to analyze the DART-MS data of six Rauwolfia species to identify the chemical markers. Thirteen and twenty three chemical markers were identified from roots and leaves which were able to discriminate among six Rauwolfia species. This method was also cross-validated for the rapid identification, authentication and quality control of Rauwolfia species to improve the efficacy of herbal drugs.

Samer M, Al-Hulu, Assistant Professor of Microbiology, has completed his PhD at the age of 29 years from Babylon University/College of Science. He has published more than 14 papers in microbiology field. Al-Hulu, has training at Ministry of Health at Laboratory of Babylon Maternity and Children Hospital. Now working at Al-Qasim Green University/College of Food Science.

Biosimilars are \"similar but not the same\" to the original biologic innovator product. Some of challenges that effect on biosimilars development such as Verification of similarity, not having of Unique name, findings of biosimilars guidelines, safety, labeling of biosimilar and regulatory framework for assessing of biosimlar. Development of Biosimilars achieved by finding of Unique name, Using of powerful analytical tools for biosimilars comparability to the reference, Quality by Design studies, Using of statistical method for reducing of simple size of biosimalir clinical trail and post-marketing surveillance studies to establish biosimilar safety, and single use bioprocessing (single use bioreactor) for biosimilar production for meeting their clinical manufactures. Pharmacovigilance programs is important for establishing clinical databases. A new generations of biosimilars such as biobetters , monoclonal antibody biosimilars having more acceptability and approved.

A current global trend in the development of biomedical and pharmaceutical chemistry is the use of computerized analytical technologies like COBAC, typically followed by the comparison with the action of the known alalytes using QSAR / QSPR methods. The clinicians are interested not in the results of the primary measurements, but in representative results of the physiological and biochemical tests compared with the preceding drugs and their analogs. In this case a direct conversion of numerical data into the QSAR / QSPR descriptor values with their subsequent transformation to the qualimetric results is necessary, which is described in this paper.

• Molecular medicine is a broad field, where physical, chemical, biological and medical techniques are used to describe molecular structures and mechanisms, identify fundamental molecular and genetic errors of disease, and to develop molecular interventions to correct them. The field of molecular medicine is often referred to as "tomorrow's medicine". It aims to provide a molecular understanding of how normal cellular processes change, fail or are destroyed by disease. Molecular Biology has proved to be a rich source of new therapeutic agents in the last three decades. Recombinant proteins continue to be developed as successful drugs that principally target extracellular proteins such as cytokines and cell-surface receptors. Protein drugs are almost always injected. Bioinformatic data can now be used to identify new intracellular target proteins and investigate the networks of interactions that the target proteins participate in. It is becoming increasingly possible to model the surfaces of target proteins and use this information to model the interaction of low molecular weight, orally available drugs and even design drugs from scratch. The completed Human Genome Project revolutionised the ways that we can consider human diseases. Single gene defects that cause rare genetic disorders took man-centuries to discover only twenty years ago. Now, because of next generation sequencing (NGS), single, novel gene defects can sometimes be identified in individual patients with only man-weeks of effort. It will soon be economically plausible to sequence all of an individual's genes in the clinic. Common diseases, though, are not caused by single gene defects. Many clearly involve the interaction of many susceptibility genes with the environment. An important part of the environment is the microbiome, the collective of microorganisms that inhabit an individual human. These organisms have strong interactions, many beneficial, with the immune system of the host and are fundamental to the understanding of common inflammatory diseases. It is now relatively simply to determine the composition of a microbiome, again by NGS. Changes that do not alter DNA sequence, known as epigenetic changes, can modulate the activity of genes too. Genes can be regulated by micro-RNA transcripts. All of these changes can increasingly be analysed by dedicated NGS methods that will be used in clinics of the future to investigate common diseases and to identify the multiple defects that drive individual patients' cancers. Our course aims to give you insights into all of these new developments and training in how to be a modern biomedical researcher.