FALL 2011 VOLUME 4 ISSUE 3

NEW User Presentations from The Bioprocessing Summit Workshop

Sriram Kumaraswamy, Ph. D., Senior Product Manager, ForteBio

ForteBio's Second Annual Workshop on Label-free Assays for Bioprocessing, Ligand Binding and Antibody Characterization was held on August 25, in conjunction with CHI's The Bioprocessing Summit conference in Boston, MA. The user workshop featured distinguished scientists from the industry as speakers who addressed new topics in bioprocess development using the Octet platform, including IgG quantitation for cell line development, detection of protein aggregates in crude supernatants, monitoring of product quality during upstream and downstream development and the latest tips and tricks for acquiring the highest quality label-free kinetic data.

The following is the list of speakers, the topics they presented and brief excerpts from their abstracts. A copy of their presentations is available for download by registered users of the fortebio.com website. Please visit the LEARNING menu at fortebio.com for the presentations.

	Characterization of the Antibody Quantitation Assay with the Octet RED System Amy Bass — Senior Research Associate II, Takeda Inc., San Francisco, CA "We have incorporated ForteBio's antibody quantitation assay, using Protein A biosensors with Octet RED, into Takeda San Francisco's Cell Line Development Platform. Results show that this assay is very specific for human antibodies showing no matrix effects from our various cell culture media. In addition, we see high precision when comparing numerous variability parameters. Titer values obtained from the Octet RED system show high correlation to Protein A HPLC values. The antibody quantitation assay is used in our cell line development platform at the 96 well, fed-batch shake flask and bioreactor stages to streamline the evaluation of clones."
	Tips and Tricks for Label Free Kinetic Analysis of Protein-Protein and Protein-Particle Interactions Islay Campbell — Field Application Scientist, ForteBio "Kinetic analysis of protein-protein and protein-particle interactions is a key application for real time, label free systems such as the Octet family of instruments. In this talk, tips and tricks for obtaining the best possible data are presented, including optimization of target immobilization and analyte binding, along with regeneration strategies (if required). The Octet data analysis software options are discussed, giving details of the analysis models and their applicability to data interpretation."
	Binding Rate Screen — A High-Throughput Assay in Soluble Lysate for Prioritizing Protein Expression Constructs Jiamin Yu — Senior Scientist, diaDexus (previously at Novartis) "Identification of constructs suitable for the recombinant protein production pipeline is a bottleneck for structural genomics efforts. A novel high-throughput approach, Binding Rate Screen, that can alleviate this bottleneck by screening crude lysates, has been developed on the Octet platform using the hexahistidine (His(6)) tag as a reporter. The constructs with the highest binding rates also exhibit high expression of soluble monomeric protein. Constructs expressing variations of the target protein can be prioritized on a time scale of minutes, providing a times savings of 10 – 100 fold."
	The Three A's of Biologicals are Activity, Activity, Activity: Case Study of Guiding Process Development towards Biological Specific Activity through Bioactivity and In Vitro Kinetic Assays Oren Beske — Vice President, Aragen Bioscience "This case study presents an example of a protein therapeutic expressed in mammalian cells that lost significant bioactivity when adapted to an industrial CHO production platform. Further investigation using various bioassays revealed that the reduction in specific activity was due to a fraction of the protein being expressed by the cells in an inactive form. Alternative purification processes were developed to identify and remove the inactive protein species in the preparation. Using the Octet system, an in vitro binding assay was rapidly developed and kinetic analysis of the interaction revealed that the inactive form of the product possessed a faster dissociation constant from the target, resulting in a reduced affinity constant."

List of Recent Publications Citing the Octet System

A Highly Conserved Neutralizing Epitope on Group 2 Influenza A Viruses
DC Ekiert, et. al. Science, 2011, 33 (6044), 843–850.

Biosensor-based Small Molecule Fragment Screening with Biolayer Interferometry
CA Wartchow; F Podlaski; S Li; K Rowan; X Zhang; D Mark; K-S Huang. Journal of Computer-aided Molecular Design, 2011, 25 (7), 669–676.

In vitro Analyses of the Dysregulated R206H ALK2 Kinase-FKBP12 Interaction Associated with Heterotopic Ossification in FOP
JC Groppe; J Wu; E. M Shore; FS Kaplan. Cells Tissues Organs, 2011, 194, 291–295.

Label Free Inhibitor Screening of Hepatitis C Virus (HCV) NS5B Viral Protein Using RNA Oligonucleotide
C Roh; SE Kim; S Jo. Sensors, 2011, 11, 6685–6696.

Detection of Deoxynivalenol using Biolayer Interferometry
CM Maragos. Mycotoxin Research, 2011, 27, 157–165.

A TPO Receptor Agonist, ALXN4100TPO, Mitigates Radiation-Induced Lethality and Stimulates Hematopoiesis in CD2F1 Mice
M Satyamitra. et. al. Radiation Research, 2011,175 (6), 746–758.

Mutations in the G-H Loop Region of Ephrin-B2 can Enhance Nipah Virus Binding and Infection
J Yuan; G Marsh; D Khetawat; C Broder; L Wang; Z Shi. Journal of General Virology, 2011, 92 (9), 2142–2152.

Challenges in Developing Bioanalytical Assays for Characterization of Antibody Drug Conjugates
JP Stephan; KR Kozak; WT Wong. Bioanalysis, 2011, 3 (6), 677–700.

Formation of Raloxifene Homo-dimer in CYP3A4, Evidence for Multi-substrate Binding in a Single Catalytically Competent P450 Active Site
JA Davis; RJ Greene; S Han; DA Rock; LC Wienkers. Archives of Biochemistry and Biophysics, 2011, 513 (2), 110–118.

Comparison of an Antibody Capture and a Cell Capture Ligand-Binding Assay to Quantify a Monoclonal Therapeutic in Serum
F Spriggs; P Aggarwal; S Deng; C Soderstrom; C Donovan. Bioanalysis, 2011, 3 (6), 605–611.

Design of a Reversible Biotin Analog and Applications in Protein Labeling, Detection, and Isolation
L Ying; BP Branchaud. Chemical Communications, 2011, 47 (30), 8593–8595.

Divergent Activities Of Osteogenic BMP2, And Tenogenic BMP12 And BMP13 Independent Of Receptor Binding Affinities
SP Berasi, et. al. Growth Factors, 2011, 29 (4), 128–139.

Development and Characterization of APRIL Antagonistic Monoclonal Antibodies for Treatment of B-Cell Lymphomas
M Guadagnoli, et. al. Blood, 2011, 117, 6856–6865.

Role of Iron and Sodium Citrate in Animal Protein-Free CHO Cell Culture Medium on Cell Growth and Monoclonal Antibody Production
Y Bai, et. al. Biotechnology Progress, 2011, 27 (1), 209–219.

Integrin Binding Human Antibody Constant Domains - Probing the C-terminal Structural Loops for Grafting the RGD Motif
MW Traxlmayr, et. al. Journal of Biotechnology, 2011, 155(2), 193–202.

Review of Transducer Principles for Label-Free Biomolecular Interaction Analysis
M Nirschl; F Reuter; J Voros. Biosensors, 2011, 1, 70–92.

For a complete list of references by year, visit www.fortebio.com/references.html.