Host Cell Proteins (HCPs) have negative impacts on the stability, safety, and efficacy of recombinant protein drugs. Therefore, it is crucial to ensure the purity of the final drug and the consistency of the purification process, which requires the use of qualified HCP ELISA kits with broad reactivity. HCP ELISA kits must meet two criteria: possessing broad reactivity and passing methodological validation. The Cygnus HCP ELISA Kit is the industry's gold standard. The following content details the correct experimental operation and methodological validation process of the Cygnus HCP ELISA Kit to help you efficiently master the key aspects of the experiment.

Proper plate washing operation is essential for reducing variability and eliminating background interference in ELISA experiments. If you are using automated plate washing equipment or the variability of your experimental results is higher than expected, we hope the following article can help you optimize the experimental process and ensure the experimental results return to normal. How to Resolve ELISA Detection Result Drift with Correct Plate Washing Methods?

HCP ELISA detection methods must undergo methodological validation in accordance with regulatory-approved experimental approaches (such as ICH or FDA guidelines) to ensure they can generate accurate results. To obtain accurate HCP ELISA detection results, essentially, it is necessary to ensure that the concentration of HCPs in the sample is higher than the Limit of Quantitation (LOQ) of the kit, and at the same time, the detection antibodies for HCPs are in excess. To meet these two requirements, it is imperative to perform dilution linearity validation on the test samples (in-process samples and final drug bulk solutions). Therefore, dilution linearity is one of the key experiments for verifying the specificity and accuracy of HCP ELISA methods. HCP ELISA Method Validation - Dilution Linearity

During the use of ELISA kits, ensuring the accuracy of results is of utmost importance. To effectively identify the compatibility between sample formulations and kit components, the spike-and-recovery test, a critical validation experiment, becomes particularly essential. It is not only an important tool for evaluating the accuracy of ELISA kits but also a reliable means to meet the methodological validation requirements for commercial ELISA kits set forth by regulatory authorities and pharmacopoeias (such as ICH or FDA guidelines).HCP ELISA Method Validation - Spike & Recovery

A blank control refers to a negative control set in the experimental design, aimed at monitoring non-specific background signals and providing a reference for baseline absorbance values. In Enzyme-Linked Immunosorbent Assay (ELISA), the introduction of a blank control can effectively identify and subtract background noise from sample readings, thereby improving the accuracy of detection results. However, in multi-analyte ELISA assays, this standardization method may not be applicable due to their inherent characteristic of high background signals. The following article will explore in detail why the use of blank controls is generally not recommended in Cygnus ELISA assays. How to Set Up Blank Controls When Using Cygnus ELISA Kits?

ELISA is a method recommended by pharmacopoeias worldwide for detecting residual HCPs in biological products, and it can determine the total amount of HCPs. Moreover, ELISA is a classic immunological detection method with simple operation. However, ELISA also has limitations in detecting HCPs. For example, it is impossible to know which specific HCPs are present in the sample from ELISA results, or which HCPs the antibodies used for detection react with. Therefore, regulatory authorities require that when using ELISA to detect the residual content of HCPs during the process, it is not only necessary to prove the accuracy, precision, sensitivity, etc., of the kit through methodological validation but also to demonstrate the broad reactivity of the HCP antibodies used, i.e., HCP antibody coverage. The AAE-MS method is used to identify information about high-risk HCPs in the final drug, thereby ensuring that these HCPs can be detected. After completing all the above methodological validation steps, this immunoassay method can be used for in-process sample monitoring and batch release of products. This decision-making process is helpful for the development and validation of universal or process-specific HCP ELISA methods, as it demonstrates how to combine orthogonal AAE and MS methods and use data to determine which type of detection method is more suitable for your project. Antibody Affinity Extraction (AAE?): A New Chapter in Innovating Host Cell Protein (HCPs) Analysis