Major characteristics of a protein are determined by its amino acid sequence. The next levels are secondary, tertiary, and quaternary structures. Notably, protein features can be modified by a variety of posttranslational modifications including glycosylations. Approximately 50 - 60% of human proteins get some kind of glycosylation usually by the addition of N- or O-linked glycans. Glycosylation cannot only affect the structure of proteins, but also their biological activity, potency, serum half-life, pharmacokinetics, pharmacodynamics, and immunogenicity.

Therapeutic proteins represent the fastest growing market of the pharmaceutical industry and there is a tremendous rush by many companies worldwide to develop Biosimilar versions of innovator products. The type of cell line used for the production of therapeutic proteins, and defined cell culture conditions have a considerable influence on the glycosylation pattern, which in turn can directly affect product quality, safety and efficacy. Carbohydrates like terminal galactose residues, bisecting glcnac and core fucose have a critical impact on mab mediated effector functions such as Antibody-Dependent Cellular Cytotoxicity (ADCC). Mannosylated glycans and sialic acid N-Acetylneuraminic Acid (NANA) can impact PK, and lower levels of galactose reduce Complement-Dependent Cytotoxicity (CDC) activity. For production of therapeutic proteins, analysis of glycosylation patterns is thus of utmost importance.

As an example, rituximab makes it clear that a biosimilar should never be developed without knowing the status of the glycosylation pattern in comparison to the originator molecule. One biosimilar in development showed higher receptor affinity and higher ADCC activity, and therefore EMA has advised the applicant to adjust the manufacturing process. After thorough analysis, the primary amino acid sequence of the biosimilar was shown to be identical, and secondary and tertiary structures of the proteins were indistinguishable. However, proportions of some glycosylations were slightly different. A modified manufacturing process finally directed the oligosaccharide composition within the variability of the originator.

Standard techniques for analysis of glycoproteins are based on high performance liquid chromatography, mass spectrometry, and capillary electrophoresis. The application of lectins as a class of molecules that can specifically bind carbohydrate-protein structures has evolved in the last years in combination with microarrays as a promising additional tool.

Lectin microarrays were first reported in 2005 and are prepared by immobilizing various lectins on a solid surface. The microarray procedure is based on an evanescent-field fluorescence-detection principle, which allows sensitive, real-time observation of multiple lectin-carbohydrate interactions under equilibrium conditions. Analytes including glycoproteins, whole cells, or bacteria are labelled with a fluorescent dye or antibody before loading onto a commercially available lectin microarray containing up to 45 specific lectins.

The usefulness of lectin microarray technology is demonstrated with several examples. In one of these, the glycosylation pattern of the mab MB311 expressed in SP2.0 cells was compared to the mab MB314 expressed in a plant expression system to generate a de-fucosylated version of MB311. The fucose indicative lectin-binding patterns correlated with the respective ADCC activities.

Such data and available literature show convincing evidence that lectin microarrays can be used for screening and evaluating different glycan patterns of therapeutic glycoproteins.

In this talk, I will take you along on my exciting journey from this first attempt all the way to developing biosimilar monoclonal antibodies. It will shed light on how much the attitude of the European regulators changed from then to now.

The requirements were inflated due to the little experience both on the side of the sponsor and the regulators. After the first products were approved and first guidelines were issued the situation calmed and requirements were based on science and evidence. Then followed the desperate attempts of the originator companies to discredit the biosimilar movement. All sorts of arguments were presented to delay and prevent the dawn of Biosimilars. The originators fought on legal grounds and countered with semi-scientific argumentation which culminated in a final push to force Biosimilars developers to do Phase III interchangeability studies.

Finally, all these efforts will have been in vain, hundreds of millions of dollars will have been wasted. But the Biosimilars will prevail. The originators will finally go back to what they are supposed to do, innovate !

Biologics that target autoimmune responses and inflammatory injury pathways have a marked beneficial impact on the management of many chronic diseases, including rheumatoid arthritis, psoriasis, ankylosing spondylitis and inflammatory bowel disease.

Crohn disease (CD) and ulcerative colitis (UC) belong to chronic inflammatory bowel diseases, which are induced by autoimmune processes. While CD is characterized by over-activity of Th1, ILC1, and MAIT cells, UC is mediated by exaggerated activities of Th2 and ILC2 cells and cytokines they produce.  Knowledge of the pathogenesis enabled a rational therapy based mostly on biologics and small molecules. 

The use of biologic therapies for the treatment of IBD is rapidly expanding, owing to the good efficacy and safety profiles of these drugs, and the better understanding of the initial targets of altered immune regulation and activity.

The major targets of most biologic therapies are cytokines, B cells, and co-stimulation molecules. Anti-cytokines include anti-tumor necrosis factor (TNF)-α, anti-interleukin (IL)-1, and anti-IL-6 molecules. B-cell depletion includes use of anti-CD20 antibodies and B cell receptor (BCR) modulation by the B-lymphocyte stimulator (blys). 

TNF is the principal proinflammatory cytokine in both diseases (CD and UC). Anti-TNF monoclonal antibodies, mostly infliximab or adalimumab were therefore introduced to their treatment.  Approximately 50-70 % of CD and more than 33 % of UC patients respond to primary treatment only, which resulted in the development of other biologics and small molecules. Out of them, monoclonal antibodies targeting adhesive molecules (vedolizumab, etrolizumab) and p40 chains shared by IL12 and IL23 (ustekinumab) have been already in clinical practice. There are also other small molecules in clinical trials: mongersen, tafacitinib, and ozanimod. Mongersen supports immunosuppressive activity of tgfβ; it has been tried for the treatment of CD. Tofacitinib inhibits activity of JAK kinases; it was shown to be effective in UC management.  Ozanimod interferes with migrations of activated T cells to the site of inflammation and is a promising drug for the UC treatment.

In 1973 the American Spinal Injury Association made the International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI). In this clinical review our patient was classified after the vertebral fixation surgery with a ASIA-A scale injury after suffering a fracture and luxation at T-12-L1, having total spinal cord section (Fig.A). Based on the research made by Sergei Paylian, phd on animal models [1] and the safety use of allogeneic mscs demonstrated on multiple animal models applications [11,12], we decided to apply a experimental translational medical protocol based the research and the previous outcomes obtained by Hamid and macewan [8,9] and decided to customize it exclusively to our patient based on the clinical evidence and personalizing the therapy on evidence. The medical team designed an ambulatory method utilizing a C-arm to apply the allogeneic mscs in situ and using a intrathecal (subdural) catheter using a slow pump release system for the rest of the biological material with an optimum tolerance and minor side effects (mild fever, miyalgias and headache) on the first 48hrs hour after application. The experimental use of mrna Bioquantine® was well tolerated with its purified form (intra and extra-oocyte liquid phases of electroporated oocytes [6]) showing to be well tolerated by the patient without any anaphylactic reaction. The current clinical report is meant to demonstrate the beneficial changes with the use of Bioquantine® and its administration in a patient with a severe SCI offering a possible optional therapy and potential neuroregeneration in this clinical condition. Mesenchymal stem cells (mscs) are ideal for cell-based therapy in various inflammatory diseases because of their immunosuppressive and tissue repair properties. Moreover, their immunosuppressive properties and low immunogenicity contribute to a reduced or weakened immune response elicited by the implantation of allogeneic mscs compared with other cell types, allogeneic mscs are a promising option because of their low immunogenicity and immunosuppressive and tissue repair capabilities. The positive findings throughout the evolution of our protocol for spinal cord injury with the obtained results at this stage is a promising scientific based and evidence based medicine protocol that can be offered in the near future as an option for severe SCI patients. The functionality of the restoresensor® surescan® by providing the electric stimulation fortifies the medical outcome and has given the patient the confidence to perform his physical rehabilitation with more energy for a longer time by the increase or decrease of the intensity of it according to the type of exercise regulated by the control-battery he handles. At this date, after 8 intrathecal applications of allogeneic mscs and Bioquantine® in situ combined together we have got the following outcomes: an improvement in sensitivity, strength in striated muscle and smooth muscle connection by increased muscle mass (Fig.B) and sphincter control, at 23 months after the first regenerative therapy and 12 months after the placement of restoresensor® the patient is showing an evident improvement on his therapy of physical rehabilitation (legs movement and control of them) having the following movements reported by the physical therapist: a)hip: adduction and external rotation, extension, abduction, internal rotation; b)knee: flexion; c)toe: MP and IP extension, also reporting an easier and functional crawling forward and backwards, and since 3 months ago the patient is capable to stand on his knees for 2 or more minutes without any support (Fig. C) and taking small steps on his knees forward and backwards for the first time in his process, showing a progressively important functionality on both limbs, voluntary movement at both feet and an increase in sensory perception. Conclusion: As we are probing in the case, combinatorial biologics can be used safely with other electronic disposals and bring a major benefit to SCI patients. With this unique combination we can give an option to those No Option patients and create many more to improve patients quality of life.

Protein-based therapies hold an enormous potential for treating many terminal diseases. Nevertheless, the lack of universal technological approaches that enable development of protein formulations with the targeted attributes significantly impedes clinical translation of these advanced therapies. This presentation will overview the use of droplet based microfluidic technology for developing protein formulations with pre-programmed functional characteristics, including size and internal morphology, encapsulation efficiency, and protein release profile.

The prevalence of breast cancer (BMD) in the world in general and in Ukraine is steadily increasing. Epidemiological, experimental and clinical studies have shown that metabolic disturbances associated with body mass index (BMI)> 30 kg / m2 increase the risk of occurrence and worsen the clinical course of breast cancer. Thus, in patients with obesity, a decrease in the sensitivity of the tumor to systemic antitumor therapy, an increase in the frequency of postoperative complications and a decrease in the rates of general and non-recurrent survival.

The aim of the study was to improve the results of neoadjuvant systemic antitumor therapy in breast cancer patients with abdominal obesity (BMI greater than 30 kg / m2) by administering levocarnitine in combination with NSAT for the correction of metabolic disorders as the main pathogenetic part of obesity.

The high development achieved by the biopharmaceutical industry in the world has allowed count in the present with innovative medicines for the treatment of chronic, life threatening and severely debilitating diseases. Recombinant proteins and monoclonal antibodies represent a great percent within the medicinal products used for the treatment of diseases such as cancer, arthritis, cardiac dysfunctions and AIDS.

Access to therapies based on the use of biological products has been limited, particularly in the third world countries, as a result of the high costs stipulated by the pharmaceutical companies that developed and patented these products. The expiration of patents and data protection for many of these products in the coming years opens the opportunity to other manufacturers to produce and market, with acceptable prices, new versions of the innovator, facilitating the access to them, which in turn translates into greater benefit for our patients.

Cuba has a biopharmaceutical industry capable of responding to the increasingly rising needs of its National Health System and it is qualified to develop new products and obtain other, for examples the biosimilars. These industry might provide so much the domestic market as that of other countries of the region. In consequence with this situation, establishing the guidelines guaranteeing the quality, safety and efficacy biosimilars products is a challenge for the National Regulatory Agency (Center for State Control on the Quality of Drugs; CECMED).

CECMED has faced during the latest years the challenge of regulating a national industry with a great scientific and high innovative potential. The methodology to elaborate guidelines for this type of products was mainly based on the revision of the legal and regulatory international framework and  the evaluation of the current and prospective developmental stage of the technology in. A first draft of the regulatory document was compared to guidance drafts issued by other regulatory agencies (such as OMS and EMEA) and extensively reviewed by CECMED specialists as well as by experts of the local biotech industry. As a result, a regulation project was elaborated, and its applicability tested while the assessment of an application for scientific advisory, related biosimilars product. Finally, the regulation was approved, being applicable for ¨Known Biological Product, term adopt in Cuba for the biosimilars products.