HuProt™ Content and Quality Assurance

Newly Re-sequenced Content and Expression

The HuProt™ human proteome microarray (v3.1) contains >20,000 unique, newly re-sequenced and individually purified human proteins representing 16,152 genes (~81% of the human proteome; see figure, right) and 124 unique mouse gene symbols.  Recombinant proteins are expressed in yeast (S. cerevisiae), purified and printed on glass slides in duplicate, along with control proteins. 

Custom selected protein panels can also be printed, processed and analysed at Arrayjet.  Please contact us for more information. 

HuProt™ v3.1 content can be searched by clicking Here .
 

Figure: HuProt arrays contain recombinant proteins expressed in yeast, individually purified and printed on glass slides in duplicates.  

 

Production and Quality Assurance

Arrayjet Inkjet Technology powers the high-throughput manufacturing of HuProt™ Arrays.

After expression, the N-terminal GST- and His6-tagged proteins are purified and printed on glass slides in duplicate, along with control proteins (GST, BSA, Histones, IgG, etc.).  Slides are barcoded for tracking/archiving.  Each microarray batch is routinely evaluated by GST staining to ensure printing quality and content.  Flexibility is built-in, meaning the HuProt™ human proteome microarray is available on two types of glass surfaces: PATH™ and SuperEpoxy 2™. 

Focused arrays can be designed to feature biomarkers identified within the HuProt™ discovery phase.  Efficient (up to 2x7) slide formats can be configured for a cost-effective and rapid validation phase.

Arrayjet's high throughput bio-printing platform, Ultra Marathon II, is used by CDI Labs to manufacture HuProt™ with significant performance advantages over contact print methods such as:

• Precise spot morphology 
• Bigger batch sizes for larger cohort analysis
• Rapid production of custom configured arrays
• Reduced inter- and intra-array variability
• Greater data reproducibility 

 

Figure: High quality content and manufacturing using Arrayjet Ultra Marathon II bio-printer, ensures reproducibility and reduces variability: (A & B) Patient serum sample run in duplicate on two separate HuProt arrays. (C & D) Patient sample from the same blood draw, serum vs plasma run on two separate HuProt arrays. Note: data points are fluorescent intensity value of auto-antibodies in patient serum bound to individual spotted proteins in the HuProt array.