DNA Removal By Depth Filters in Bioprocesses

Question By: Ashish Vishwakarma

Ans.: Dear Ashish, I would like to tell you that this is a Downstream Processing from Bioprocess purification Process in which gene coding for therapeutic proteins are inserted into host cell and fused with myeloma cell to produce hybridoma cell. these cell are grow in to the fermentation vessels in order to produce the desired therapeutics proteins.
Therapeutics proteins are transported in to culture medium by the host cell and purification begin with separation of the cell mass from the protein suspended in the culture medium. during these separation process host cell DNA can be release from dead or lysed cell.

eg. Zeta Plus depth filters
Process: DNA in final product dosage forms should not exceed 100 pg per dose. Host cell DNA can contaminate the culture fluid as a result of release by lysed or dead cells. In some cell culture clarification systems, centrifugation or tangential flow filtration (TFF) may be used as a primary clarification step. Both of these technologies are shear intensive and can increase cell lysis, resulting in increased host cell DNA contamination. The presence of contaminating DNA early in a process creates the possibility for carry through during downstream purification. It is therefore important to remove as much DNA as possible during the early stages of purification.

Fact: Zeta PlusTM depth filters, contain cellulose, filter aid, and a resin that imparts a positive charge to the filter surfaces. Zeta Plus depth filters are commonly used for initial clarification of cell culture broth and cell lysates. These filters contain significant voids volume (hence the name depth filter) that allow for accumulation of cellular debris. The filter structure is a series of interconnecting pore pathways able to retain cellular debris by mechanical entrapment. In addition to debris removal by mechanical retention, Zeta Plus filters are able to remove particles smaller than their pore size. This capability is based on electrokinetic attraction of negatively charged particles by the positively charged filter surfaces. It was anticipated that DNA, which is a polyanionic molecule, would also be removed by Zeta Plus filters.


CONCLUSION:

The removal of DNA from cell culture broths and lysates may be required to accomplish the CBER-recommended concentration of DNA impurity of <100 pg/dose in therapeutic dosage forms.

In many purification processes, it is desirable to remove impurities as early as possible from the process. 3M Purificationfs Zeta Plus depth filter medium has
been extensively used in upstream processes to clarify cell culture broths and cell lysates and serves as an effective means to reduce contaminating levels of DNA.

Reference: www.bioprocessintl.com/.../archive/.../BPI_A_100807AR23_O_103770a.pdf

Note: Dear ashish please patch your comment for further clarification

Bioprocess Engineering


Bioprocess Engineers work at the frontiers of biological and engineering sciences to “Bring Engineering To Life” through the conversion of biological materials into other forms needed by mankind. Application areas commonly associated with bioprocess engineering include the production of biofuels, design and operation of fermentation systems, development of food processing systems, application and testing of product separation technologies, design of instrumentation to monitor and control biological processes, and many more. Like other engineering disciplines, bioprocess engineers are trained in the application of engineering sciences and problem solving techniques. What separates them from other engineers is their understanding of how every engineering field relates to living materials. With the rapid growth of biologically based technologies, bioprocess engineers are uniquely qualified to solve the problems of today and tomorrow. The demand for bio-process engineers continues to grow. They provide a bridge between the research lab and the economic, large-scale implementation of biotechnologies and food production systems.
Professional Organizations
Professional organizations bioprocess engineers can become affiliates of include:
Institute of Biological Engineering
American Society of Agricultural Engineers
Institute of Food Technology
American Institute of Chemical Engineers
International Society for Pharmaceutical EngineeringOpportunities in Bioprocessing
With its agricultural heritage and high technology industrial base, North Carolina and the U.S. provide a wide range of opportunities for the Bioprocess Engineer. Biotechnology- based applications are poised to leave the research lab and enter commercial production.
Over the next 20 years in North Carolina these industries are expected to generate up to $15 billion in annual sales and employee up to 100,000 North Carolinians. North Carolina Agribusiness ranks third nationally in terms of net farm income and provides 22.5 percent of the total gross state product. The U.S. food industry is currently employing 20,000 engineers from all disciplines and exceeds $400 billion in annual sales, and represents over 25% of all nondurable goods produced. Each of these provides potential career opportunities for the well-trained engineer with the knowledge, interest, skills and commitment to be a part of this diverse and increasingly important field.
Bioprocess engineers are employed in:
U.S. food industry
International corporations
Biotechnology companies
Government agencies
Private consulting firms
Agricultural commodity processors

Bioprocessing- Scop & 0pportunity

Bioprocessing- scope & opportunity
Bioprocessing makes use of microorganisms, cells in culture or enzymes to manufacture products. Humans have been using such processes for baking bread, making cheese and fermenting alcoholic beverages since prehistoric times. Advances in recombinant DNA technology allow the production of an enormous variety of protein-based therapeutics that are having a profound impact on the quality of life for severely ill patients. Bioprocessing is also key to several emerging industries and technologies, including the production of renewable biofuels such as ethanol and biodiesel, therapeutic stem cells, gene therapy vectors, and new vaccines. The Food and Drug Administration imposes stringent regulations on bioprocessing in the biotech industry; this highly regulated environment has an enormous impact on plant operations and product manufacture
Students in the Bioprocessing focus area will obtain a thorough understanding of industrial processes from early-stage development through large-scale manufacture.
Career OptionsBioprocess EngineeringProcess DevelopmentManufacturing OperationsQuality Assurance/ComplianceRegulatory Affairs (cGMP)Project ManagementEnvironmental RemediationFood TechnologyTherapeutic Stem CellsDevelopment and Manufacture of Gene Therapy Vectors and VaccinesDevelopment and Production of Renewable BiofuelsNew Enterprise Development
Topics in the CurriculumBioprocess Engineering FundamentalsBioprocess Design and EconomicsUnit Operations and Process Flow DiagramsBioprocess Operations ManagementBioreactor Design and OperationsMicrobial FermentationMammalian and Stem Cell CultureBioseparationsBioprocess Development & Scale-upCurrent Good Manufacturing Practices (cGMPs)Standard Operating Procedures (SOPs)Recombinant Protein ProductionProtein Recovery, Purification and AnalysisBeer and Wine FermentationViral Vaccines and Gene Therapy Vector ProductionRenewable Biofuels Production