Anaquant HCP analysis I Protein characterisation I Protein analysis
Throughout time, our expertise in identifying and quantifying Host Cell Protein (HCP) has become more and more accurate. We have developed a unique technology capable of detecting and quantifying HCPs, thus enhancing any bioprocess. Our HCPprofiler solution has a very efficient way to detect and quantify HCPs in any sort of matrices from a no complex matrix to a highly complex one.
This article will present to you how the ANAQUANT HCPprofiler method has been tested and how our experts have demonstrated its efficiency in quantify HCPs in all bioprocess matrices.
How is HCP-PROFILER an innovative solution?
The ANAQUANT HCPprofiler kit is composed of both an innovative standard and a custom data processing software. The standard consists of a hydro-soluble bead containing peptides at different concentrations to provide an internal calibration curve. In this process, one bead of a standard is added to a sample of a variable matrix.
Then, the sample analysis and identification are processed thanks to the HCPprofiler software. It allows to convert MS signal into amount of protein, in an easy-to-use fashion. The collected data is very simple to read and enables you to compare samples or any other sort of results to provide an internal report based on your needs.
Our purpose of our work was to solve matrices complexity issues and to ensure quantification of HCPs whatever the DownStream Process step (DSP). Within the course of our experiments, we successfully demonstrated that HCPprofiler standard is stable and reproducible whatever the considered matrices.
Dive into our method and results
We have analyzed several heterogeneous kinds of samples with HCP-PROFILER in order to assess the repeatability of the standard. The goal was to ensure that the standard was as efficient in a no complex matrix as it could be in a highly complex matrix. Thus, we tested samples from yeast, vaccines, mRNA, cell culture, etc.
A total of 14 samples of matrices with different complexities have been tested:
Samples were processed according to a simple guideline. Firstly, a sample of a no complex matrix to a highly complex matrix was taken and submitted to digestion, reduction, alkylation or desalting. Secondly, the HCPprofiler standard was added to the sample to generate a calibration curve.
The standard used in this process is accurately calibrated using the READYBEADS™ technology. Afterwards, samples were analyzed with the last generation of high-resolution MS instruments. Finally, data were processed with the easy-to-use HCPprofiler software. 
To illustrate and assess sample heterogeneity, total protein amount was estimated in each one. As a result, the total protein amount was encompassed between 8 ng and 42 µg into the samples, while the number of identified proteins was found between few dozens and several thousands in the different samples. 
It is important to notice that after MS acquisition and data processing, calibration curves were automatically obtained with HCPprofiler software. The calibration curves were compared based on several criteria:
- The slope;
- The intercept;
- The regression coefficient.
For each one of these criteria, deviation between samples were found in less than 6%, which leads us to think that, whatever the considered sample, calibration curves are indeed repeatable. Since slopes are conserved whatever the matrix, the results we obtained confirmed the reproducibility capabilities of HCP-PROFILER.
Our conclusion and remarks
After spending months studying repeatability across different matrices with HCPprofiler, we can now confirm that our innovative standard and custom data processing software are a well-adapted solution for quantification of Host Cells Proteins. HCPprofiler is able to ensure reproducible calibration curves, whatever the matrix complexity.
This standard is able to solve the reproducibility problems and biases user-induced while it ensures stability over several months. It definitely appears as a valuable tool to ensure accuracy and reproducibility in quantitative studies.
Should you like to learn more about HCPprofiler or any of our innovative solutions, feel free to contact us.