Arrayjet flying inkjet biochip spotting technology for the production of human proteome chips HuProt

Arrayjet flying inkjet biochip spotting technology for the production of the world's highest throughput human proteome chip HuProt

Arrayjet AdvanceTM production service increases chip yield while reducing sample consumption

Figure 1: After the installation of the Ultra Marathon II in Baltimore, USA, the customer is very satisfied with the instrument (left).
Ultra Marathon II plus Jetmax environmental control system for spotting at very low temperatures (right).

Case
CDI Labs is an American proteomics company that previously used a low-throughput contact pinning platform. They experienced frequent production delays, reduced production, large batch-to-batch variations, and sample loss. They lack a technology platform to produce high-throughput, high-quality protein chips. As demand continues to increase, CDI faces the pressure to effectively commercialize their products and reduce rising equipment maintenance costs.

Arrayjet Adance chip spotting service
Arrayjet's Adance chip spotting service started in 2011 and is very effective in supporting CDI's projects. This direct-to-customer chip service provides customers with direct support from Arrayjet's 75 years of comprehensive biochip experience to optimize, transfer and commercialize their protein chips.

Experimental optimization
A portion of the human protein in the human protein library is spotted onto the epoxy silane (Figure 2) and nitrocellulose membrane via Arrayjet's Ultra Marathon II flying inkjet biochip spotting platform. The entire spotting environment is controlled by the Jetmax environmental control system. 4 ° C. The sample volume at each last point is optimized by testing various spot volumes. Among the 14 micromatrix repeats, the sample point circular morphology yield rate is greater than 99%. The microarray is used to obtain the expected expression profile.

Figure 2: Experimental optimization on an epoxy silane chip

Batch sampling
More protein samples from the CDI Human Protein Library were spotted onto 200 sheets of epoxy silane and nitrocellulose slides for further analysis of spotted repetitive spotting patterns (Figure 3). Arrayjet's Jetguard ensures minimal sample evaporation during long-term spotting.

Figure 3: A panel of protein samples purified from the human protein library was spotted onto 200 identical Grace Bio-lab PATH nitrocellulose membranes.

High-density spotting <br> Confirmed by functional protein experiments, we successfully carried out the methodological transfer from pin point to Arrayje flight inkjet spotting. We further evaluated the ability of the Ultra Marathon II flight inkjet spotting platform for high-density spotting. Samples of the CDI Human Protein Library were subjected to a high-density hexagonal matrix dot to evaluate the reproducibility of on-chip and inter-chip spotting, as well as background signals, on different spotted matrices. The results show that the chip has a very good matrix with no dot overlap.

Whole Human Proteome Chip Preparation <br> More than 19,000 GST fusion proteins were purified from the CDI Human Protein Library and repeatedly spotted onto 500 sheets of 200 pl samples per point (Figure 4). The criteria for successful experimentation are as follows:
• >97% of samples need to be spotted onto the chip • Number of round dots >90%
• CV <20% in chip and chip
• Positive and correct functional test results

Figure 4: CDI's complete human protein library sample was spotted onto a Grace Bio-Labs PATH® protein chip slide.

Conclusion <br> Through the successful transfer of methodologies, CDI is currently able to produce the world's largest human proteome chip, producing 100 versions of the 3.1 version of the HuProt chip in one batch. Purchasing the Ultra Marathon II flight inkjet spotting platform allows CDI to prepare proteomic chips and other custom-made chips. With these chips, customers can perform molecular interaction testing of tens of thousands of proteins using a minimal amount of clinical samples. Arrayjet Advance chip services and non-contact piezoelectric spotting technology significantly increase the productivity of large-scale, high-quality protein chips. The environmental control unit not only guarantees the perfect sample point morphology and matrix, but also protects the natural conformation of the protein, and ultimately guarantees the consistency of the experimental results.

“CDI purchases the Ultra Marathon II flight inkjet spotting platform for the development and production of specific chips for HuProt protein chips and hybridoma cell screening projects. CDI is planning to use this new technology platform to prepare monoclonal antibody chips in the near future. And membrane protein chips. We chose Arrayjet because we needed to achieve a 5-fold increase in flux, and it is better to have an instrument to achieve this. Arrayjet's high-tech, precise and user-friendly design is a distinct advantage. The advantage of an Arrayjet company structure is that it also provides internal chip services through Arrayjet Advance. We can deepen our trust in the platform through testing of several projects over several months. This platform can enhance us in many aspects. Currently operating."
Dr. Ignacio Pino, CEO, CDI Laboratories"

Arrayjet's Jetspyder sample introduction device effectively reduces any cross-contamination between protein samples and the quality of our protein chips has improved significantly. Previously, we were able to produce only 150 quality-qualified chips in one batch, a flux that did not meet the needs of scale production and current and future growth. Arrayjet's technology platform is very attractive for us to quickly and efficiently produce 1000 chips. ”

Dr. Heng Zhu, Professor, Johns Hopkins School of Medicine

Health Monitor

Health Monitor (Health Care Monitor) is based on the principle of pulse pressure method, and based on the established theory, the model is divided into compartments, and a series of calculation formulas are deduced by linear correlation algorithm. After these formulas are corrected by nonlinear compensation and clinical experience parameters, a set of reflecting cardiac function, vascular condition, blood state and microcirculation function can be obtained.