1. What techniques do you currently use for the analysis of samples?
Various types of ELISA assays (sandwich assays and competitive assays) with colorimetric, fluorescence and chemiluminescence detections are used to analyze the samples

2. What other technologies do you currently use?
Currently, QPS' Immunoanalytical group has four colorimetric readers, a fluorescence reader, a luminescence reader, a electrochemiluminescence (ECL) reader, and two Bio-Plex machines (Luminex technology).

Mulitplex bead based assays have been validated using the Bio-Plex (Luminex technology). These assays can help determine the amount of mulitple analytes in a single sample.

3. What program do you use to capture the data from your experiments?
We use the Softmax pro 3.1.2 to produce capture and analyze the ELISA data and to calculate standard curve from our colorimetric assays and the Softmax pro version 4.8 for the fluorescence assays.

In addition we use the Softmax pro for Lmax version 1.1L to analyze the data from the luminescence reader.

4. Do you have a GLP compliant version of software for data management and report writing?
QPS uses Watson LIMS v 6.4 for validation and sample analysis projects.

5. How many ELISA methods do you develop and validate in a year? What is ratio of method development vs. method transfer?
Between 2005 and 2007, QPS' Immunoanalytcal group developed and fully validated about 70 methods, and performed about 15 cross-species and/or cross-matrix partial validations. The ratio of method development to method transfer is approximately 4:1.

6. What is the scope of your method development for GLP studies?
QPS will construct the ELISA assays based on the available antibody, and perform antibody conjugation as necessary. The initial assay conditions will be developed through a feasibility study.

The method will have to demonstrate the following before going into validation:
 ►Minimum required dilution
 ►Precision and accuracy of full assay range
 ►Analyte selectivity test in matrix (minimum 6 lots)
 ►Linearity of dilution (or parallelism if endogenous)
 ►Stabilities of focus

7. Do you perform in-house validations?
We can validate a Sponsor's ELISA method or prepare our own ELISA method for your specific analyte(s).

8. What is the process to validate an ELISA assay? What are your validation criteria for ELISA assay?
The ELISA assays are validated by multiple runs usually involve more than one analyst. A typical validation includes 6 runs for precision and accuracy, room temperature and freeze/thaw stability, matrix selectivity, dilution test, hook effect, and long-term frozen stability.

The scope of the full or partial validation which follows the latest Crystal City Bioanalytical Validation guideline, and the AAPS Journal 2007; 9 (1), E30-E42 article "Workshop/Conference Report - Quantitative Bioanalytical Methods Validation and Implementation: Best Practices for Chromatographic and Ligand Binding Assays" is defined below. A validation report will be issued which will be suitable for regulatory filing.

The full validation will demonstrate:
 ►Precision and accuracy (3-day quality controls with minimum 4 concentration levels, including LLOQ)
 ►Dilution test (1 concentration level, three dilution factors; n > 2)
 ►Matrix Selectivity test (LLOQ level; six individual lots)
 ►Sample stability:
 ►Long term frozen stability (at about one month, minimum of 2 concentration levels @ -20oC or below; n = 5). The concentrations will lie within the validation assay range.
 ►Room temperature stability (minimum 4-hour cycle; 2 concentration levels; n = 5)
 ►Freeze/thaw stability (one 24-hour cycle, and two 12-hour cycle; 2 concentration levels; n = 5)
 ►Post preparative re-injection reproducibility (minimum of 3 concentration levels in reconstitution solvent in duplicates at room temperature)

9. Do you run your standard curves and QC with matrix samples?
Yes, most ELISA assays for PK analysis are run with calibration standards and QCs prepared in the intended matrix. For assays that endogenous level is expected, calibration standards may be prepared in buffer, while the QCs can be a combination of buffer and matrix based preparation for different levels.

10. What are some of the matrices you have worked on?
Mostly serum and plasma (EDTA>Heparin>Citrate) from mouse, rat, rabbit, monkey, and human. We have also a small percentage of urine samples. Occasionally we work with peripheral blood mononuclear cell (PBMC) or platelet lysate, as well as tumor tissues. Recently we were assigned to the analysis work that involves monkey milk. For cellular assays we have performed whole blood incubation and assays using established cell-lines.

11. How many protocols and samples do you run in a year? What is the ratio of clinical to preclinical/nonclinical samples?
Between 2005 and 2007, QPS' Immunoanalytical group worked on approximately 80 protocol-driven studies, which equates to approximately 122,000 samples. The ratio of clinical to preclinical/nonclinical samples averages about 60/40 over the years. The exact ratio depends on the development cycle of our sponsors.

12. Do you have the capability to run assays with radioactive samples?
Currently we do not encourage studies using radioactivity.

13. How many scientists work in your particular group?
23 scientists currently work in the Immunoanalytical-Protein Biomarker group and will expand throughout the year.