Tag: <span>testing</span>

Cell substrates are used in various stages of viral vaccine manufacturing, as in the isolation, selection, and propagation of the virus seed or virus vector stock, as well as for the amplification of the virus to produce the final vaccine product. The various stages of cell substrate use, including cell banking, are shown in a generic manufacturing scheme in Figure 1. Traditionally, viral vaccines have been produced in animal tissues, primary cell cultures, and cell lines that either have a finite life span, such as normal diploid cells, or a theoretically infinite life span, as achieved with continuous or immortalized neoplastic cells. The cell substrates used in viral vaccines currently licensed in the US are listed in Table 1…

Manufacturing Viral Vectors

Contamination by adventitious agents (bacteria, fungi, mycoplasma, and viruses) represents potential safety risks for biologics produced in mammalian cells. Bacterial and fungal contaminations are usually easy to detect in culture medium due to changes in pH and visual indicators such as color and opacity. Mycoplasma contamination has been detected in 15‚Äď35% of cell lines deposited in some cell culture collection. This is because mycoplasma contaminations often cause little changes that can be readily detected by visual inspection. However, bacterial, fungal, and mycoplasma contamination can be more effectively controlled than viral contamination by careful screening of initial parental cell banks, proper environmental monitoring, along with ongoing testing‚Ķ

Cell & Tissue Banking

Recombinant DNA-transduced cellular products encounter the product development and regulatory issues of both gene therapy and cellular therapy products. The characterization of recombinant DNA-transduced cellular products remains highly challenging for both sponsors and regulatory agencies. The regulatory concerns and product testing for such cellular products are similar to those for all biologicals. These concerns include the demonstration of product safety, identity, purity, and potency; the control of the manufacturing process to ensure the consistency of product manufacturing under a proper quality control program; and the demonstration of reproducibility and consistency of product lots by means of defined product lot release testing criteria…

Cell & Gene Therapy Regulatory

Psoriasis is a chronic, inflammatory disease of the skin that is estimated to affect 2-3% of the U.S. population. The estimated annual outlay for treating the disease has ranged from $1.6 billion to $3.2 billion, and the cost to individuals with the disease is far higher than the monetary costs. Psoriasis is characterized by excessive keratinocyte proliferation, leading to a significant thickening of the epidermis, expansion of epidermal rete pegs into papillary dermal space and abnormalities in the differentiation process. Clinically, one sees red, raised, scaly lesions that can occur over any part of the body. The etiology of the disease is complex and not well understood. T-cells are almost certainly involved in the initiation and maintenance of psoriatic lesions. Although the triggering event is immunological, alterations in keratinocyte function also appear to be important to the overall pathophysiology…

Biologics Production

The Vmax™ technique has been used extensively to estimate filter area requirements for normal flow filtration (NFF) processes in biopharmaceutical applications. The benefits that this technique presents over conventional flow decay methods are the speed of testing, reduced volume requirements for evaluation, and competitive testing of varying filter types/sizes ‚ÄĒ all of which present an optimized filter screening strategy and preliminary estimate of optimized filter size requirements. Filter size or filtration area requirements derived using the Vmax technique consist of contributions from both capacity and flow-time aspects of the filtration process. This article examines the relative contributions of these terms to overall filter sizing vis-√†-vis the ease of fluid filterability‚Ķ

Biologics Production Uncategorized

Manufacturers must demonstrate, with a very high degree of assurance, that biopharmaceutical products derived from mammalian cells or from human plasma are safe and free of viral contamination. Viruses can be physically removed from most proteins using filtration. Often air diffusion is used as a nondestructive test to ensure that a process filter is installed properly and free of defects that can compromise virus retention. In this article, theoretical models were used to relate air and liquid flow rates through integral and defective filters. The effect of defect diameter and defect density on the virus retentive ability of a filter was also modeled…

Manufacturing

Although biological products are being licensed at a fairly steady pace, the cost to develop each product can be incredibly high, and far too many products with very little chance of success are entering clinical trials. The cost of developing a biological product is now estimated to be as high as $1.7 billion. This is truly a staggering figure that would seem to prevent all but the strongest company from attempting such a gamble. However, this number includes the cost of all the products that didn‚Äôt make it through pre-clinical development, or which entered clinical trials and failed for any number of reasons…

Biologics Production Regulatory

The United States Pharmacopeia (USP) is a 184-year-old organization that has been in the forefront of technology since its inception. From publishing a manual about how to prepare therapeutic potions, USP has evolved into a compendium of standards and information on manufactured pharmaceutical products, with more than 4,000 monographs covering drug substances and biologics, and their dosage forms, excipients, and nutritional supplements. It is not surprising that the USP initiative in cell and gene therapy and tissue engineering has closely followed the emergence of these technologies…

Cell & Gene Therapy Regulatory Risk Analysis and Management

Validating the safety of biological preparations requires thorough testing for contamination by adventitious agents. Utilizing mammalian cell cultures to produce recombinant proteins as biopharmaceuticals requires testing for viral contamination. The polymerase chain reaction (PCR) may be employed to specifically detect the presence of viral DNA or RNA with great sensitivity. PCR assays are particularly useful for the qualification of recombinant cell banks. Regulatory agencies recommend that mammalian cell banks be tested for a variety of possible human viral contaminants. In most cases the cells used to produce the cell bank have been previously analyzed for viral contamination. The use of PCR for the detection of viruses in the final banked cells can alleviate the need for difficult, costly, and time-consuming infectivity assays. In some cases the relevant viruses cannot be cultured, eliminating the ability to perform infectivity assays. The PCR assay can provide a sensitive and specific method for detection of viral contamination when standard infectivity assays are unsatisfactory…

Cell & Tissue Banking

The first HIV-based lentiviral vector to be used in humans, VRX496, is currently being tested in Phase I clinical trials (initiated in January 2003). With each new therapeutic comes the need to establish quality control (QC) testing designed specifically for that product. The QC testing for VRX496 was developed in accordance with the Code of Federal Regulations (CFR) 21 for pharmaceutical and bulk chemical GMPs, Points to Consider in the Characterization of Cell Lines Used to Produce Biologicals (1993) from the Center for Biologics Evaluation and Research (CBER) at FDA, and the United States Pharmacopeia (USP) 1046 for Cell and Gene Therapy Products. This report describes the QC testing of lot VRX496V02-009 of our clinical grade vector, which is being used in an ongoing clinical trial evaluating the first lentiviral gene therapy vector in humans. All assays are performed according to established standard operating procedures (SOPs) and in accordance with the principles of cGMP regulations…

Cell & Gene Therapy Viral Vectors