KRIBIOLISA™ CHO HCP ELISA

SKU: KBBP03 Category:

₹35,000.00

Immunoassay for the quantitative measurement of CHO Host Cell Proteins (Chinese Hamster Ovaries) as a contaminant in various biological preparations


Availability: 3 – 4 Weeks | Pack Size: 1 x 96 wells



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Description

Introduction:
A variety of proteins which are used as therapeutic agents in humans and animals are produced through recombinant expression in Chinese Hamster Ovary (CHO) cells. The manufacturing and purification process of these products tends to leave the potential for contamination by Host Cell Proteins (HCPs) from CHO cells, which may result in adverse toxic or immunological reactions that ultimately affect the efficacy of the therapeutic agent. The simple, objective and semi-quantitative ELISA is a highly sensitive method that aids in purification process development, process control, quality control and product release testing optimally.

Intended Use:
This generic KRIBIOLISA CHO HCP kit is intended in determining the presence of Chinese Hamster Ovary Host Cell Proteins contamination in various products that are manufactured through recombinant expression in CHO cells. The kit has been validated successfully for testing of in process and final product HCPs in variety of products regardless of growth and purification process.

Principle:
This assay is based on the Sandwich ELISA procedure. Samples containing CHO HCPs are reacted with already coated affinity purified capture anti-CHO HCP antibody. Following an incubation period unbound components are removed by a washing step. Then the anti-CHO HCP: HRP is added. This immunological reaction results in formation of a sandwich complex of solid phase antibody-HCP-enzyme labeled antibody. The wells are again washed to remove any unbound reactants. The TMB substrate is then added to the well. The amount of hydrolyzed substrate is read on a microtiter plate reader and it is directly proportional to the concentration of CHO HCPs present.

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Additional information

Sample Type

Cell Culture Supernatant and Other Biological Preparations
Calibration Range

0.312 – 20 ng/ml
Sensitivity

0.188 ng/ml
Detection Method

Colorimetric
ELISA Type

Direct Sandwich Assay
Regulatory Status

For Research Use Only
Storage Temperature

Store the unopened product at 2-8 Deg C. Do not use past expiration date.

Citations

Protocol

Assay Procedure:
It is strongly recommended that all Standards and Samples be run in duplicates or triplicates. A standard curve is required for each assay.
1. Add 100 ul prepared Standards and diluted Samples to respective wells.
2. Cover the plate with a sealer and incubate for 90 minutes at 37?C.
3. Aspirate and wash plate 4 times with diluted Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe of any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
4. Pipette 100 ul Biotinylated CHO HCP Antibody Working Solution to all wells.
5. Cover the plate with a sealer and incubate for 60 minutes at 37?C.
6. Aspirate and wash as per Step (4) above.
7. Pipette 100 ul Streptavidin:HRP Conjugate Working Solution to all wells. Mix well.
8. Cover the plate with a sealer and incubate for 30 minutes at 37?C.
9. Aspirate and wash as per Step (4) above.
10. Pipette 100 ul TMB Substrate in all the wells.
11. Incubate the plate at 37?C for 20 minutes. DO NOT SHAKE or else it may result in higher backgrounds and worse precision. Positive wells should turn bluish in color.
12. Pipette 100 ul of Stop Solution to all wells. The wells should turn from blue to yellow in color.
13. Read the absorbance at 450 nm with a microplate within 10-15 minutes after addition of Stop solution.

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