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The Establishment and Stability of the Peripheral Blood Mononuclear Cell Donor Bank for Immunoassay Validation

by Janet L. Lathey, PhD, Scott Gregory, Mike Ewell, and Scott Hickman
Volume 5, Issue 1 (Spring 2006)

The enzyme-linked immunospot (ELISpot) assay is one of the most useful techniques for the immunological monitoring of vaccine trials and has increasing application as a measure of specific T-cell activation. Recently, we developed, optimized, and validated a customized ELISpot kit for the detection of interferon gamma (IFNγ) positive cells. The precision of the ELISpot was good and it varied over the range of the assay values, independent of the stimulus. Here we describe the development of a library of donors with characterized responses to the CEF peptide pool: cytomegalovirus (CMV), Epstein-Barr virus (EBV), and influenza (Flu); pool of 32 peptides which can be used as controls for IFNγ ELISpot and multiple immune monitoring assay validations for use in clinical trials...

Citation:
Zhang C. The Establishment and Stability of the Peripheral Blood Mononuclear Cell Donor Bank for Immunoassay Validation.
BioProcess J, 2006; 5(1): 64-68.

 
Protein, Peptide, and Amino Acid Analysis with Corona CAD: A New Universal HPLC Detector

by Darwin Asa
Volume 5, Issue 1 (Spring 2006)

The quantitative and qualitative analysis of proteins and their amino acid sequence composition is a critical operation in many researc laboratories and operations. Much protein analysis is performed using high-pressure liquid chromatography (HPLC) with ultraviolet (UV) or fluorescence detection. Although these methods are robust and widely used, certai issues limit their utility. For example, some proteins or peptides may have a poor UV response and can be difficult to detect. Direct comparisons of protein levels when quantitated directly by UV can alson be problematic due to differences in extinction coefficients of various proteins...

Citation:
Asa D. Protein, Peptide, and Amino Acid Analysis with Corona CAD: A New Universal HPLC Detector.
BioProcess J, 2006; 5(1): 69-73.

 
An Overview of the United States Pharmacopeia's Recent Activities in the Areas of Vaccines, Virology, and Biological Standardization

by Tina S. Morris, PhD
Volume 4, Issue 4 (July/August 2005)

The United States Pharmacopeia (USP) is a not-for-profit nongovernmental organization founded in 1820 that develops public standards for drug substances and products; these standards are enforceable by FDA and have been adopted by many nations around the world. USP General Chapters provide industrial and academic researchers alike with crucial guidance, particularly in areas where there is a regulatory void. A good recent example is the proposed USP general information chapter...

Citation:
Morris TS. An Overview of the United States Pharmacopeia's Recent Activities in the Areas of Vaccines, Virology, and Biological Standardization.
BioProcess J, 2005; 4(4): 46-50.

 
Large-Scale Chromatographic Purification of an Attenuated Chimeric Poliovirus

by Tom Ouellette, Steven L. Giardina, PhD, Earl Nelson, Judith Poiley-Nelson, PhD, Trevor Broadt, and Sibylle Herzer, PhD
Volume 4, Issue 2 (March/April 2005)

Poliovirus is a small (28-30 nm diameter) non-enveloped RNA virus belonging to the family Picornaviridae. The ability of poliovirus to cross the blood-brain barrier and its natural infectivity of central nervous system (CNS) tissue via the CD155 receptor, found exclusively in primates, has promoted the investigation of an attenuated poliovirus for the treatment of malignant gliomas. However, use of the virus in clinical testing is limited due to low yields obtained from conventional purification methodologies...

Citation:
Ouellette T, Giardina SL, Nelson E, Poiley-Nelson J, Broadt T, Herzer S. Large-Scale Chromatographic Purification of an Attenuated Chimeric Poliovirus.
BioProcess J, 2005; 4(2): 31-38.

 
A Novel Human Expression System for Generating Glycoprotein with Higher Activity, Fully Human Glycosolation, and Optimized Sialylation

by Hans Baumeister, PhD, Ute Schöber, Marion Schlangstedt, PhD, Renate Stahn, PhD, and Steffen Goletz, PhD
Volume 4, Issue 2 (March/April 2005)

Today concentrated efforts are underway to improve the bioactivity of therapeutic proteins with the aim of reducing: (i) the number and concentration of the applied doses of the therapeutic protein, (ii) undesired side effects, and (iii) the cost of a therapy. A very promising strategy is to optimise the glycosolation of these biotherapeutics. A novel expression platform, GlycoExpress™, has been developed to produce proteins with fully human glycosolation, optimised sialylation, and improved bioactivity...

Citation:
Baumeister H, Schöber U, Schlangstedt M, Stahn R, Goletz S. A Novel Human Expression System for Generating Glycoprotein with Higher Activity, Fully Human Glycosolation, and Optimized Sialylation.
BioProcess J, 2005; 4(2): 45-50.

 
Report from the Lentivirus Vector Working Group: Issues for Developing Assays and Reference Materials for Detecting Replication-Competent Lentivirus in Production Lots of Lentivirus Vectors

by Veronique Kiermer, PhD, Flavia Borellini, PhD, Xiaobin Lu, Vladimir Slepushkin, PhD, Gwendolyn Binder, Boro Dropulic, PhD, Muriel Audit, PhD, Barbara Engel, MD, Kenneth Cornetta, MD, Carolyn Wilson, PhD, Dan Takefman, PhD, Yuan Zhao, PhD, and Keith Carson
Volume 4, Issue 2 (March/April 2005)

The Lentiviral Vector Reference Working Group (LVRWG) was created at the conclusion of a meeting organized by The Williamsburg BioProcessing Foundation in June 2002, in conjunction with the American Society of Gene Therapy (ASGT) annual conference. The meeting participants were gathered to evaluate the need for developing reference material to ensure comparability of lentiviral and retroviral vectors, in a similar spirit to the Adenovirus Reference Material program that had just been completed. The concensus at the conclusion of this meeting was that the diversity in the lentiviral vector field, which includes vectors derived from different parental viruses and with various designs, does not allow for identification of a single reference material that would benefit more than a single or very few investigators...

Citation:
Kiermer V, Borellini F, Lu X, Slepushkin V, Binder G, Dropulic B, Audit M, Engel B, Cornetta K, Wilson C, Takefman D, Zhao Y, Carson K. Report from the Lentivirus Vector Working Group: Issues for Developing Assays and Reference Materials for Detecting Replication-Competent Lentivirus in Production Lots of Lentivirus Vectors.
BioProcess J, 2005; 4(2): 39-42.

 
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