Continuous virus filtration: An existing technology with a promising future

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Continuous bioprocessing and process analytical technologies: A path towards quality by design

Recent success in monoclonal antibody based immuno-oncology therapeutics such as Keytruda® has initiated a revolution within the Biopharmaceutical Industry. Development efforts to support this class of molecules include single-use technology, continuous production, process analytical technology (PAT), information technology (IT), multivariate data analysis (MVDA), and efforts towards real time release testing (RTRT). Merck’s vision for a next-generation large molecule production facility aligns with these six key principles, as demonstrated by the construction of a continuous monoclonal antibody (mAb) production pilot plant named the Protein Refinery Operations lab (PROLab) within Merck Bioprocess Development. Quality-by-Design (QbD) principles are maturing for the development of standard batch-based therapeutic protein manufacturing processes. Outside of the insight gained through similar techniques applied to unit operations run in a continuous mode, similar approaches for the dependencies created by connected and continuous processes are still in their infancy. Previously, a methodology for characterizing holistic downstream process performance through real time perturbation analysis, where the response to an upstream stimulus is monitored at several unit operations simultaneously was presented to start applying QbD principles to continuous bioprocessing. Here, the authors build upon the concept of perturbation analysis by presenting case studies where advanced PAT tools have been embedded in PROLab operation. At-line and off-line process and product quality data from continuous upstream and downstream production will be presented over the course of long term perfusion cell culture with variable production rates. Specific emphasis will be placed on the performance of bioreactor cell retention and clarification trains, and post-chromatographic ultrafiltration unit operations. This process understanding can then be utilized to place PAT tools at the appropriate locations in the process to achieve product attribute control and ultimately to assure uninterrupted supply of therapeutic proteins to patient

Inactivation of Clostridium sporogenes and Geobacillus stearothermophilus spores with the use of microwave and steam sterilizers and microwave oven

Introduction. Equipment for sterilization used in medical laboratories must be absolutely effective in eliminating microorganisms and their spores. It often directly influences human health, even life. The aim of the study was to compare the effectiveness of sterilization using the steam sterilizer ASV E, microwave sterilizer EnbioJet ML1, microwave sterilizer for baby bottles and breast pumps AVENT and microwave oven. Materials and methods. Evaluation of the effectiveness of sterilization with the use of selected devices based on pressure-thermal and microwave-thermal methods was conducted, on the basis of elimination of G. stearothermophilus PCM 2104 and C. sporogenes IW 1306 spores. Results. After using the steam sterilizer, 100% inactivation of spores of both species was noted. In the case of EnbioJet ML1 sterilizer, in the test containing 106 CFU × cm–3 G. stearothermophilus spores, 1.63 × 101 CFU × cm–3 survived. The baby bottles sterilizer proved less effective. While the microwave, in the case of tests with the highest spore content, provided their inactivation only at the level of more than 70.0%. The steam sterilizer and EnbioJet ML1 sterilizer were the most effective, whereas the latter ensured a very short time of high temperature effect, which has a favorable impact on the properties of sterilized products, for example compounds decomposing in high temperature. Conclusion. Results of own, as well as other authors’ studies allow to confirm the large potential in the scope of using microwave radiation for the sterilization and disinfection of materials of various sensitivity to temperature.

Wykrywanie biofilmu in vivo na powierzchni siatki chirurgicznej - opis przypadku

We present the case of a 51-year-old male who having undergone abdominal hernia repair with the use of synthetic implant, presented bacterial biofilm which had formed secondary to surgical wound infection. The presence of biofilm on the implant hindered the treatment of the infection. Full recovery was possible only after the implant’s removal. The presence of bacterial biofilm was confirmed in vivo on randomly selected fragments of the mesh implant.Przedstawiono przypadek 51-letniego mężczyzny poddanego operacji przepukliny brzusznej z użyciem wszczepu syntetycznego, u którego w wyniku zakażenia miejsca operowanego na powierzchni siatki chirurgicznej powstał biofilm bakteryjny. Obecność biofilmu na implantacie była przyczyną oporności zakażenia na leczenie. Wyleczenie uzyskano po całkowitym usunięciu implantatu. Na losowo wybranych fragmentach siatki chirurgicznej potwierdzono in vivo obecność biofilmu bakteryjnego

Highly automated bioburden-free continuous manufacturing biologics GMP operations: How to get there?

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BEZOBJAWOWE ZAKAŻENIE SIATKI CHIRURGICZNEJ – OPIS PRZYPADKU

Infection involving a surgical implant is one of the most serious complications associated with the use of biomaterials in hernia surgery. Implant infection may manifest clinically in a number of ways. The authors present a case of asymptomatic infection of a mesh implant which had been used to repair a paraumbilical hernia. The infection was diagnosed accidentally during surgery for recurrence. The presence of a biofilm on the surface of the old implant was confirmed by a quantitative method based on 2,3,5-triphenyltetrazolium chloride(TTC) and by scanning electron microscopy (SEM). The biofilm served to protect the microorganisms from the activity of the patient’s immune system, resulting in an asymptomatic clinical course of the infection. It is the authors’ opinion that all implants which are removed during surgery for recurrent hernias should be routinely evaluated for the presence of microorganisms even if no apparent signs of infection can be observed. The TTC method should be included in the diagnostic tools in order to limit the percentage of false negative results.Zakażenie obejmujące implantat jest jednym z najpoważniejszych powikłań towarzyszących stosowaniu biomateriałów w chirurgii przepuklin. Zakażenie implantatu może mieć różny przebieg kliniczny. W pracy przedstawiono przypadek bezobjawowego zakażenia siatki chirurgicznej zastosowanej do zaopatrzenia przepukliny okołopępkowej. Zakażenie zostało rozpoznane przypadkowo podczas operacji z powodu nawrotu przepukliny. Badania metodą jakościową z użyciem chlorku 2,3,5-trójfenylotetrazoliowego (TTC), metodą ilościową oraz z użyciem skaningowego mikroskopu elektronowego wykazały obecność biofilmu bakteryjnego na powierzchni implantatu zastosowanego do pierwotnego zaopatrzenia przepukliny. Jego obecność na powierzchni implantatu uchroniła drobnoustroje przez działaniem układu odpornościowego pacjenta i była odpowiedzialna za bezobjawowy przebieg zakażenia biomateriału. Zdaniem autorów wszystkie implantaty usuwane podczas operacji z powodu nawrotu przepukliny należy poddać badaniu mikrobiologicznemu, nawet gdy nie stwierdza się makroskopowych cech zakażenia. Do badań diagnostycznych powinna zostać włączona metoda redukcji TTC, co pozwala ograniczyć liczbę wyników fałszywie ujemnych