Rockland Immunochemicals, Inc. recently launched the BioQuantiProTM CHO-HCP ELISA Kit (KJA-4001) for the sensitive detection of host cell protein impurities in bioprocessing. The kit was specifically designed with Rockland’s novel, validated antibodies to detect HCP contaminants and to measure analytes from crude cellular harvests, cell culture lysates, and process intermediates. The BioQuantiProTM CHO-HCP ELISA Kit boasts the highest coverage of generic CHO-HCP kits, low variability, and, most importantly, the data to back it all up.
This kit meets industry standards for specificity and sensitivity and is specifically designed for standard to high throughput applications and can be easily adapted for automation. The key component of this kit is a broad-coverage antibody produced against a spectrum of host cell contaminants including both low abundancy and high abundancy proteins, as well as poorly immunogenic and highly immunogenic proteins.
• Coverage: 89% total coverage across LMW & HMW proteins
• LLD: 2 ng/mL
• LLQ: 4 ng/mL
• Range: 2-200 ng/mL
• Intra variability: <10%
• Inter variability: <15%
• CHO HCP Antibody coated 96 Well Strip-Plate
• HCP Kit Sample Buffer
• CHO HCP Protein Standard
• HCP Kit Wash Buffer (10X)
• CHO HCP Detection Antibody (300X)
• HCP Kit TMB Buffer
• HCP Kit Stop Buffer
• Plate sealer
The CHO-HCP ELISA Kit (KJA-4001) is a sandwich, enzyme-linked immunosorbent assay. The impurities in process-derived samples containing CHO-K1 proteins are captured and detected in a microtiter-type plate by using pre-coated, purified, rabbit polyclonal, CHO-HCP-specific antibodies. The immunological reaction after the addition of TMB results in a chromogenic change that can be measured photometrically at 450 nm with reference wavelength at 630–650 nm. The O.D. in the wells is directly proportional to the amount of HCP concentration and correlates to the amount of HCP present in the original sample. The HCP concentration can be calculated based on the calibration curve generated with the provided CHO-HCP standard.
Rockland’s CHO-HCP ELISA Kit uses purified rabbit antibodies generated against CHO-K1 cell lysate that were derived from cells grown in chemically defined media. The antibodies developed for this kit were validated for broad coverage of CHO-HCP samples using two-dimensional (2D) Western blots. Appropriate coverage across a 2D-blot is demonstrated using 2 criteria:
1.Percent total coverage: determined by identifying the number of detected protein species compared to the number of existing species shown on a corresponding 2D SDS-PAGE.
2.Quadrant analysis: demonstrate the proper detection of proteins having diverse sizes and isoelectric points.
Well-developed HCP reagents show broad coverage that includes commonly difficult to detect proteins in the lower molecular weight regions (LMW). The quadrants are defined as:
Quadrant 1 (Q1): HMW proteins (above 50 kDa) in low pH region (below pH 6.5)
Quadrant 2 (Q2): HMW proteins in high pH region (above pH 6.5)
Quadrant 3 (Q3): LMW proteins (below 50 kDa) in low pH region
Quadrant 4 (Q4): LMW proteins in high pH region
The CHO-HCP ELISA Kit reagents demonstrated broad coverage and identified difficult-to-detect low molecular weight (LMW) proteins in Q3 and Q4, in addition to high molecular weight (HMW) proteins in Q1 and Q2. This data indicates a robust reagent suitable for use in an HCP ELISA kit.