{"product_id":"holistic-bioprocess-engineering-of-antibody-fragment-secreting-bacillus-megaterium-von-florian-david","title":"Holistic bioprocess engineering of antibody fragment secreting Bacillus megaterium","description":"\u003cp\u003eAntibodies and antibody fragments are most important tools for therapeutic and diagnostic\u003c\/p\u003e\u003cp\u003eapplications. An increasing demand of these high potential drugs makes less cost\u003c\/p\u003e\u003cp\u003eintensive production systems most desirable. Compared to mammalian cell culture\u003c\/p\u003e\u003cp\u003emicrobial production platforms are beneficial with regards to reaching high production\u003c\/p\u003e\u003cp\u003etiters, scale up approaches, regulatory aspects and reduced costs of goods. Amongst\u003c\/p\u003e\u003cp\u003ethese production systems those that are having the ability of an additional product\u003c\/p\u003e\u003cp\u003esecretion are most advantageous as the downstream processing costs are decisively\u003c\/p\u003e\u003cp\u003ereduced.\u003c\/p\u003e\u003cp\u003e\u003c\/p\u003e\u003cp\u003eIn this work Bacillus megaterium, as a Gram positive model organism, was used to\u003c\/p\u003e\u003cp\u003eextensively study the production and secretion of the antibody fragment D.13 scFv. For\u003c\/p\u003e\u003cp\u003ethe bioprocess optimization a holistic approach was followed. First the aim was to\u003c\/p\u003e\u003cp\u003eestablish a high productive defined cultivation medium. Different media components like\u003c\/p\u003e\u003cp\u003ecarbon sources, metal ions and ammonium concentrations were screened throughout\u003c\/p\u003e\u003cp\u003evarious cultivation platforms ranging from micro titer plates to shaking flasks. Statistical\u003c\/p\u003e\u003cp\u003edesign of experiments and a genetic algorithm approach were used to establish an\u003c\/p\u003e\u003cp\u003eappropriate defined high production medium. As a second step the process was transferred\u003c\/p\u003e\u003cp\u003eto the bioreactor scale of several liter cultivation volume. An optimal bioprocess\u003c\/p\u003e\u003cp\u003estrategy based on alternating growth and starvation phases was established to gain high\u003c\/p\u003e\u003cp\u003eproduct titers of antibody fragment D1.3 scFv. As a final step an up-scale to a 100 L\u003c\/p\u003e\u003cp\u003ebioreactor was done accounting for an advanced process control and considering ¿good\u003c\/p\u003e\u003cp\u003emanufacturing practice¿ guidelines. The advanced bioprocess monitoring tool of flow\u003c\/p\u003e\u003cp\u003ecytometry was used to gain deeper insights on microbial physiology at single cell level\u003c\/p\u003e\u003cp\u003eregarding cell viability, cell integrity and production intensity. This knowledge was used for\u003c\/p\u003e\u003cp\u003ea sophisticated cell physiology based bioprocess development and optimization. Furthermore\u003c\/p\u003e\u003cp\u003eculture heterogeneities were measured and characterized for B. megaterium\u003c\/p\u003e\u003cp\u003eproducing antibody fragment D1.3 scFv under controlled bioreactor conditions. To obtain\u003c\/p\u003e\u003cp\u003eadditional information about the regulatory processes occurring inside the cell on gene\u003c\/p\u003e\u003cp\u003eexpression level a transcriptome analysis was performed comparing cells with an\u003c\/p\u003e\u003cp\u003eincreased production and secretion status to less producing and non-producing cells.\u003c\/p\u003e\u003cp\u003eThese cutting-edge technologies of flow cytometry and transcriptome analysis revealed\u003c\/p\u003e\u003cp\u003epossible bottlenecks of the overall bioprocess¿ performance and product secretion of\u003c\/p\u003e\u003cp\u003eantibody fragments with B. megaterium as production platform and form a robust basis for\u003c\/p\u003e\u003cp\u003erational strain optimization and advanced process designs.\u003c\/p\u003e\u003cdiv class=\"aw-variant-hidden-subtitle-div\" id=\"aw-variant-subtitle-9783954041152\"\u003e\u003ch3\u003e\u003c\/h3\u003e\u003c\/div\u003e","brand":"Autorenwelt Shop","offers":[{"title":"Softcover - 9783954041152","offer_id":40129827766365,"sku":"9783954041152","price":26.85,"currency_code":"EUR","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0940\/0622\/files\/cddd118e-4e8c-4d00-84c2-ed1e2230049e.jpg?v=1756880230","url":"https:\/\/shop.autorenwelt.de\/products\/holistic-bioprocess-engineering-of-antibody-fragment-secreting-bacillus-megaterium-von-florian-david","provider":"Autorenwelt Shop","version":"1.0","type":"link"}