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  PA 800 plus Pharmaceutical Analysis System--

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PA 800 <i>plus</i> Pharmaceutical Analysis System

Therapeutic proteins make up a rapidly growing segment of global pharmaceutical production. These complex molecules require accurate characterization of product purity, heterogeneity and identity. This includes data regarding their stability, shelf life and related manufacturing processes.

Analysts handling therapeutic proteins need:

•   Automated, qualitative and quantitative analysis
•   Simplified functionality and maximum operational efficiency
•   Robust, validated applications that can be transferred globally

The PA 800 plus Pharmaceutical Analysis System addresses these needs by providing robust analytical tools for the development and quality control of therapeutic proteins.

Automated and Quantitative Analysis of Biologics

The PA 800 plus Pharmaceutical Analysis System* was designed in collaboration with biopharmaceutical development and QC groups. This platform provides analysts with robust and easy-to-use characterization, integrating quantitative, qualitative and automated solutions for protein purity, charge isoform distribution and glycan analysis.

The PA 800 plus offers dependable, accurate determination of protein purity, heterogeneity, and identity. To create it, Beckman Coulter collaborated with biopharmaceutical development and QC groups experienced in the routine use of capillary electrophoresis for protein characterization.

Learn more about PA 800 plus applications and software.


Product Features
A Robust Solution for Demanding Research Requirements
The PA 800 plus offers dependable, accurate determination of protein purity, heterogeneity, and identity. To create it, Beckman Coulter collaborated with biopharmaceutical development and QC groups experienced in the routine use of capillary electrophoresis for protein characterization.

Automated Sample Introduction
The PA 800 plus system offers fully automated methods and extended sample-handling capability for walk-away operation. Sampling can be performed using 1.8 ml universal vials, 96-well plates and micro vials. Precision-molded polymethylpentene universal vials accommodate run buffer, sample and micro vials.

Sample Temperature Control
Sample temperature control maintains molecular stability when working with temperature-labile protein species. The sample temperature can be maintained between 4° - 60°C.

Temperature Control of the Capillary
Efficient separations in CE rely on precise regulation of the capillary temperature to manage Joule heating within the capillary. Proper temperature control plays an important role in the repeatability of SDS-gel, cIEF and carbohydrate analysis.

The PA 800 plus uses recirculating liquid coolant to provide effective heat dissipation when performing assays on the system. Capillaries are housed in patented cartridges, facilitating both temperature control and easy exchange of capillary dimensions and surfaces (US Patent #5198091; see application bulletin T1823ab). Capillary temperature can be regulated between 15° - 60°C.

Multiple Modes of Sample Introduction and Separation
The PA 800 plus offers electrokinetic, pressure and vacuum injection of samples. Additionally, injection from either end of the capillary allows both ultra-fast and high-resolution analysis. Separations can be adjusted by varying voltage, current, pressure and vacuum. The combination of voltage and pressure in the SDS-gel assay ensures the gel buffer stays free of air bubbles which can be generated from gel outgassing.

  • Photodiode array detection lets you perform high-sensitivity analyses across a broad range of wavelengths. Photodiode array detection between 190 and 600 nm allows for baseline subtraction and spectral wavelength analyses.
  • The PA 800 plus UV/Vis Detection Module provides absorbance spectroscopy in the UV-visible region. Commonly used exclusion filters at 200 nm, 214 nm, 254 nm and 280 nm are provided to increase analyte specificity.
  • The 488 nm solid state laser module provides robust fluorescence technology in a quiet, energy-efficient and compact design.

Variable Pressure and Vacuum
The PA 800 plus operates with all common rinsing protocols, regulating them with a pressure-handling capability of -5 to 100 p.s.i. Capillary conditioning is accomplished by moving specific volumes of electrolytes, gels, regenerants and cleaning solutions through the capillary. Gel buffers are quickly and efficiently pumped into the capillary.

Versatile Modular Detection Capability
Each PA 800 plus offers precise, real-time analysis for a variety of assays, because it integrates UV, photodiode array and LIF detection capabilities in one unit. UV detection is important when using photosensitive capillary surfaces. Photodiode array detection between 190 and 600 nm allows for baseline subtraction and spectral wavelength analyses. A 488 nm solid-state laser and laser-induced fluorescence (LIF) detector permits high-sensitivity analysis of labeled molecular species.

CE-MS Ready
Capillary electrophoresis separation coupled to mass spectrometry (MS) combines the high-resolution separation of CE and the high-sensitivity mass determination of MS. The PA 800 plus accommodates direct MS connection through the right-side access panel. Capillary temperature control is retained.

Applications
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SDS-Gel Application and IgG Purity and Heterogeneity Determination
The PA 800 plus Pharmaceutical Analysis System automates size separation of proteins and provides high-resolution, quantitative data, as compared to traditional labor-intensive slab gels.

The CE SDS-gel application has become the gold standard for protein purity analysis in biopharmaceutical laboratories. Denatured proteins can be reduced or left intact for separation and subsequent analysis.

Beckman Coulter’s patented replaceable SDS-gel consists of a polymer matrix that allows for:
  • Quantitative and automated separation of proteins from 10-225kD
  • Sensitivity equivalent to silver-stained gels when using LIF detection**
  • High-resolution separation capability


  • Beckman Coulter's Automated SDS-MW Analysis Chemistries are designed for the separation and sizing of protein-SDS complexes using a replaceable gel matrix. The gel is formulated to provide an effective protein sieving range of approximately 10 kDa to 225 kDa. Within this size range, the logarithm of protein molecular mass is linear with its reciprocal electrophoretic mobility, allowing the molecular weight of an unknown protein to be estimated from a standard curve of known protein sizes.

    The PA 800 plus IgG Purity and Heterogeneity Assay Kit was designed in collaboration with biopharmaceutical analysts developing and manufacturing therapeutic mAb molecules. Assay methodology involves heat denaturation of IgG in the presence of SDS, followed by size separation using high-resolution capillary gel electrophoresis technology.

    Capillary Isoelectric Focusing (cIEF)

    We have undertaken extensive development of cIEF to include optimization of the cartridge aperture, various cIEF chemical components, peptide pI standards and mobilization techniques, paving the way to enhancement of resolution and reproducibility. These system improvements have proven valuable not only in the development of a broad platform method for analysis of monoclonal antibodies (mAbs), but also in the enhancement of narrow range cIEF separation methods. Subsequent intermediate precision studies provide compelling evidence that high performance cIEF separations can be achieved throughout the pH gradient.

    In cIEF, a mixture of sample and ampholyte is introduced into a capillary and subjected to electrophoretic separation. In this process, a pH gradient through which analytes migrate to their respective pI is formed. Comprehensive optimization of multiple assay parameters has been performed.

    Successful transfer and implementation of characterization assays between laboratories is based on a method’s ability to minimize environmental and operator variability. An important indicator of the necessary robustness is intermediate precision.

    Carbohydrate Labeling and Analysis

    Glycosylation on a protein is an important post-translational modification that can affect its function, clearance and stability. The PA 800 plus Pharmaceutical Analysis System simplifies the complex process of profiling carbohydrates associated with glycoproteins. By providing specific and accurate quantitation of glycosylation levels, differences in glycoform quantities and distribution can be determined.

    The PA 800 plus Carbohydrate Labeling and Analysis Chemistries contain the reagents, buffers and separation capillaries required to label, separate and quantify oligosaccharides and monosaccharides released from glycoproteins. After release (enzymatic or chemical), sugars are labeled with a fluorophore, APTS, at the reducing termini by reductive amination. The stoichiometry of labeling is such that only one APTS molecule is attached to each molecule of oligosaccharide. These highly charged and fluorescent oligosaccharides are easily resolved in an electric field and detected by laser-induced fluorescence.


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    PA 800 plus Software – As Easy as 1, 2, 3…
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    The PA 800 plus Software includes features that allow you complete control of the PA 800 plus and P/ACE™ MDQ Systems. The software includes methods and sequence table development capability, advanced data analysis functionality, and includes the analytical tools necessary for proper integration and quantitation of data. In addition, advanced custom reporting can be done in order to output results the way you want to see them. The direct control capability allows for more direct user interaction with the system. Additionally, the software also allows for single-point control for all detectors configured on these systems.

    The PA 800 plus software quickly guides users from set-up through routine system operation. Large icons provide intuitive guidance for navigation while on-screen cues indicate system progress at a glance. Insightful Help menus and descriptive system prompts further simplify operator learning, making transfer of PA 800 plus technology to other analysts easier. Ultimately, using PA 800 plus software is as easy as 1, 2, 3.

    PA 800 plus software features include:

  • Automated sequence table and reagent calculations
  • Validated applications for SDS-gel, cIEF, and carbohydrate analysis
  • Enhanced Help menu and instructional videos
  • Technical controls enabling regulatory compliance
  • On-screen prompts for monitoring system events
  • Advanced reporting capability
  • Increased workspace
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    * The lamp in this product contains mercury. Dispose according to local laws.

    **Salas-Solano O., Tomlinson B., Du S., Parker M., Strahan A., Ma S. "Optimization and Validation of a Quantitative Capillary Electrophoresis Sodium Dodecyl Sulfate Method for Quality Control and Stability Monitoring of Monoclonal Antibodies." Analytical Chemistry 2006, 10.1021.

    For laboratory use only; not for use in diagnostic procedures.

    Additional Information:


     

     
     
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