Manuel Fuentes from the Cancer Investigation Centre (CIC), Salamanca, was lucky enough to work in collaboration with researchers at Harvard Medical School in formulating Nucleic Acid Programmable Protein Arrays (NAPPA).
By eradicating the need for recombinant protein production and purification, NAPPA delivers a highly reproducible and economical solution for biomarker and drug discovery in tumor and autoimmune pathologies.
Having produced results by high-medium density immunoassays in array format, to then pin-spot nucleic acids for translation to proteins in situ, the team used a Genetix instrument. The contact-printed arrays displayed cross-contamination, lacked reproducibility and were slow to produce.
The need for speed
The methodology driving NAPPA demands the involvement of cDNA and capture of antibodies in a master mix which has to be printed within a short time frame. The conventional instrument was deemed no longer fit for purpose due to critical success factors, as shown in Table 1.
Table 1. Critical success factors against results with Genetixprinter. Source: Arrayjet Ltd
|Critical success factor||Result with Genetix printer|
|Speed||The short timeframe in which both samples were to be deposited meant that a maximum of 20 slides could be printed in each run|
|Reproducibility||Samples must be printed homogenously to ensure equal protein content. The contact approach did not offer satisfactory reproducibility|
|Sample conservation||A minimum of 10 µg of costly sample was required to print only 20 slides|
|Reliability||Spots must be free of cross contamination|
The Arrayjet solution
Arrayjet’s non-contact inkjet technology generates spots of visibly improved morphology compared to pin-spotting systems (Figure 1). Resultingly, each spot was equivalent to a comparable quantity of protein for reliable data acquisition.
Figure 1. Homogeneity of sample distribution compared between contact pin spotter (left) and Arrayjet printer (right). Image Credit: Arrayjet Ltd
As well as being more reliable, Arrayjet’s “on-the-fly” method is also the fastest available on the market, facilitating deposition rates of up to 640 features per second. Sample volume requirements are significantly reduced, and print runs are generally automated, which gives back invaluable scientist time.
Transfer to the Arrayjet platform completely satisfied all key critical success factors, surpassing customer expectations (Table 2).
Table 2. Critical success factors addressed by Arrayjet technology. Source: Arrayjet Ltd
|Critical success factors||Arrayjet resolution||Benefit|
|Speed||“On the fly” printing||High density slides printed without compromising sample|
|Reproducibility||Instruments and components with CV values <5%||Comparable protein quantities translated on each spot|
|Sample conservation||Jetspyder™ liquid handling device||1.3 µL sample aspiration prints hundreds of slides|
|Reliability||Integrated wash station||Print head and Jetspyder™ do not contribute to contamination|
The benefits of using the Arrayjet printing platform are summarized in Table 3.
Table 3. Benefits of technology transfer to Arrayjet platform. Source: Arrayjet Ltd
|Benefit of Arrayjet technology over contact printing|
|Speed||Printing time reduced fourfold leaving scientists free to conduct research|
|Throughput||Fivefold increase in slide production|
|Sample conservation||Sample volume demands reduced tenfold|
|Cost||Significant financial savings associated with automation, printing speed and sample conservation|
Arrayjet’s high-throughput printing can be modified to accommodate a wide range of applications, including but not limited to:
- Antibody validation
- Antigen discovery
- Biomarker screening
- Epitope mapping
- Gene expression profiling
- Host-pathogen interaction screening
- Hybridoma screening
- Small molecule library screening
Arrayjet remains open to any innovative screening ideas that push its technology to its limits by offering both an in-house bioprinting service and a range of five scalable instruments.
Transfer to Arrayjet technology has had huge benefits for our cancer research and biomarker discovery. Our team can now produce 100 slides in a working day, using just 1 µg of cDNA per gene of interest. The automated process enables us to focus fully on our research knowing arrays will be printed reliably and reproducibly every time,”
Manuel Fuentes, Scientific Researcher at Cancer Investigation Centre
About Arrayjet Ltd
Arrayjet provide instruments and services to the pharma, diagnostic and life science industries. Our products use inkjet technology for precision picolitre liquid handling. Arrayjet focus on printing samples to create tools for genomic and proteomic screening, patient stratification and clinical diagnosis.
The proprietary printing technology is fully automated and delivers benefits of ease of use, precision, reproducibility, efficiency of manufacture, and total process-control.
Arrayjet’s patented technology simultaneously aspirates and prints multiple samples on-the-fly. This is a proven platform and its non-contact bioprinting is ideal for microarray and 96 well microplate manufacture; as well as bioprinting onto biosensors, biochips, MEMS devices, microfluidic devices, membrane sheets and into nanowell applications. Most substrates are compatible with the technology.
Arrayjet instruments offer the largest manufacturing batch size of up to 1000 slides, allowing over 18,400 samples to be loaded at once. The instruments are modular and scalable, enabling customers to increase capacity as their requirements grow. They combine the fastest and most reliable instrumentation on the market with the versatility to print any biological sample type onto any solid substrate.
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