Fundamental advantages vs other approaches

  1. BLAST doesn’t “kill” the immune repertoire prior to sampling it

  2. This, along with creative screening, enables the desired antibody to be selected from thousands of positives whether it be via ELISA, bead, or cell-based assays.

  3. Hybridoma, phage display, EBV transformation fall short in that neither samples the natural repertoire and in the event that any hits are attained, they go through months of work to create something that is still 1000X less effective than antibodies derived via BLAST. (see IL-6 and TNF mabs in clinical trials).

  4. Speed

  5. Antibodies to targets are seen 7 days from obtaining PBMC’s to seeing secreted, assayable antibodies where they are interrogated based on desired design criteria.

  6. No immortalization-based technology or current display technologies comes close in timing. 

  7. Once found - BLAST antibodies are clinic-ready - no in vitro somatic hypermutation, no lengthy humanization (ours takes nanoseconds).

  8. Diversity    

  9. Since BLAST leaves the repertoire intact, you have the luxury of “numbers”.  Massive antibody hits equals diverse epitope recognition.  The fact that BLAST can be performed in several species amplifies the possible diverse epitope detection.  All of this means that antibodies with rare, desirable properties have a high probability of being found.

  10. Industrialized

  11. BLAST works at the speed of thought.  It is the only antibody technology that can be pulled from the freezer, mixed with PBMC’s (fresh or frozen) and produce antibody hits in seven days.  It was designed for high throughput antibody production. 

High

low

~100

Cut off @ 12,000

Phage Display

BLAST

High chances of success?:  BLAST vs Phage Display

CD20 - comparing hit rate in mice immunized with the extra-cellular domain of CD20

  1. Splenocytes were halved - 1/2 for fusion, the other for BLAST

  2. Hybridoma yielded 12 IgG hits

  3. BLAST yielded 18,000 IgG hits

CD20: Comparison to hybridoma

Design Goals for a CD40 mab

  1. Bind CD40

  2. Cannot block CD40 ligand

  3. Must bind cynomolgus CD40

  4. Must cover as many epitopes as possible (~15)

  5. Need high affinity and moderate affinity to each epitope

  6. Looking for internalizing mab

Hits available

Chances?

Technique

BLAST vs the industry

vs

NCBvIndustry.html

Program Examples:

Many projects have design goals like this.  Most people who work with North Coast are accustomed to technologies that provide them with 100 or less hits to their targets which may be the reason for their low expectations.  With BLAST, we ask the partner to start with their desired product and we work backwards to the original screen.

Today there are four antibodies in clinical trials that were discovered with a BLAST-like procedure.  Two of these are to new targets and are fully human (found by the iSTAR platform), the other two neutralize ligands and they are from rabbits.  Currently what is known about these antibodies are:
The human derived, anti-infective mabs  are commonly found to be more potent than those antibodies made in non-human systems 
The anti-cytokine antibodies are 1000X greater affinity than the current antibodies made in both hybridoma and phage display technology

The vast majority of approved antibodies are from hybridoma efforts with only one from phage display.  While there’s hundreds more validated targets, this is good news for those who would rather invest in fast follow-ons to known validated targets.  BLAST-derived antibodies should outperform those antibodies interrogated simply because they were some of the few to survive versus BLAST where you select from the whole repertoire of sequences.

Illustrations of BLAST vs other mind-sets or approaches are hereNCBvIndustry.htmlshapeimage_27_link_0
BLAST Advantages: