Impact of contracted manufacturing organization protocols on operations in an aca demically based Current Good Manufacturing Practice facility.

Immunotherapy of cancer and other diseases is often dependent on adoptive transfer to patients of cellular products generated in Current Good Manufacturing Practice (cGMP) facilities. With the availability and approval of various cellular products for therapy, cell production facilities are experiencing unprecedented growth in demand for services. Increasingly, these services involve processing of externally generated cells for transfer to the bedside. The arrival of cells from external manufacturing facilities for processing and eventual infusion of cell therapy products into patients creates a new layer of responsibility and adds to an already demanding list of the existing procedures in academic cGMP facilities. Sponsors introduce their own requirements for the handling of cells that the laboratory must incorporate and follow. The challenges of creating additional access to cleanrooms, writing new standard operating procedures, expanding personnel training, altering pre-existing schedules and incorporating additional monitoring for safety of external products alter the balance of laboratory operations. Adjustments for accommodating externally manufactured products are numerous and varied, as each sponsor has requests that are product-specific. If cells produced by several different external manufacturers are handled by the same facility, the negative impact on the regular activities in this facility may be considerable. Here the authors provide a review of operational challenges that an academic-based laboratory faces and discuss solutions that could ameliorate the difficulties related to an increasing volume of industry-sponsored trials. The solution may be the development under the auspices of the Foundation for Accreditation of Cellular Therapy or the Food and Drug Administration of regulations that will guide the processing of products manufactured by external companies and make these regulations broadly applicable in all cGMP facilities.

Immunologic monitoring of cancer vaccine trials using the ELISPOT assay.

Cancer vaccines are designed to activate an immune response to tumor-specific or tumor-associated antigens expressed by the tumor. Cancer vaccines take many forms, including synthetic peptides, tumor cells and lysates, cell lines, and autologous antigen presenting cells like dendritic cells. The target antigens may be known, or "defined" in the vaccine, or unknown. In melanoma, more so than in other cancers, a large number of immunogenic "shared" antigens (tumor-specific or tumor-associated) have been identified. This allows for vaccination of groups of patients with the same vaccine, and also allows for testing for melanoma tumor immunity even when the vaccine does not include defined antigens. For the cancer vaccine field, the goal of a prognostic or predictive biomarker has yet to be achieved. However, the primary immunologic goal of any cancer vaccine is the induction (or amplification) of an immune response against the tumor, therefore the primary goal of immunologic monitoring in this setting, is testing for that response. In this chapter, we present standardized methodology from a central immunologic monitoring laboratory for melanoma cancer vaccine immune response assessment by the Enzyme-Linked Immunosorbant Spot (ELISPOT) assay. This assay allows for enumeration of antigen-specific cells in a plate format. We present the Interferon (IFN)-γ-producing lymphocyte assay, but the platform is easily adjusted to several cell types and several secreted molecules.