Fully human antibodies are the mainstream of therapeutic antibody development because of the increasing concern on immunogenicity and safety issues. Among the techniques that develop fully human antibodies, Human Naïve library and Phage Display have been proved* to be an effective platform to pave the way to the fully human candidates.
*Adalimumab, Belimumab, Raxibacumab, Ramucirumab, Necitumumab, Avelumab, Ranibizumab and Durvalumab are known to be developed by phage display and antibody library technology.
GenScript ProBio has successfully developed in-house Human Naïve library, which has been internally validated for multiple targets. Besides, our experienced scientists will develop multiple panning strategies, which is just right for you to get the best antibody candidate.
GenScript ProBio has build human naïve library in format of Fab, with library size of 7x1010, and it is still under expanding at present. Having delivered rich experience in dozens of internal case studies and clients projects, we’ve proved that GenScript ProBio’s library is qualified to for most targets, with average affinity reaching from 10-8 to 10-11mol/L and average binder No. ranging from 20-30.
|Technical Indicator||GenScript ProBio Library Indicator|
|Source||2200 healthy donors|
|In frame rate/ORF rate||>85%|
|CDR3 diversity||Normal distribution|
|Germline analysis||Nature frequency|
Provide diversified panning & screening strategies to ensure diversity & affinity
Solid panning, liquid panning, cell panning, combination panning, competitive panning tailored to different type of targets
Up to 2 panning strategies are used to maximize the success rate
No more than 3-round panning to retain the sequence diversity
Further binding confirmation for the unique leads are performed by soluble format
FASEBA HTP screening combined with ELISA/FACS binding and SPR affinity ranking
High quality delivery: Human antibody leads with high affinity(10-8~10-11 mol/L) generated in case studies
Fast development timeline: Starting from 2 months for fully human lead generation
Readily integrated downstream services: Affinity Maturation, Developability
Generally, the protein structure of drug target is varied in different conditions: some maintain its nature structure, but some changed during purification and expression. Taking this into consideration, the multiple panning strategies could help on getting the binder that exactly meet the requirement.
Besides, there are also differences in the efficiency and results of different panning strategies.
At GenScript ProBio, we will evaluate the properties of targets, and choose the best panning strategy accordingly, all for the best antibody candidates for our clients.
|Liquid panning||Better at presenting all possible epitope, high diversity|
|Solid panning||Simple format, suitable for most target|
|Cell panning||Generating antibodies able to recognize natural epitope presented by cells|
|Ag.biotin-cell panning||Generating antibodies able to recognize more diversified epitope and natural epitope presented by cells|
|Ag-cell panning||Generating antibodies able to recognize natural epitope presented by cells|
|Competitive panning||Eliminate antibodies targeting irrelevant epitope, acquiring antibodies specific to desired epitope|
Till Dec. 2019, GenScript ProBio has successfully delivered dozens of clients’ projects. Below are results of part of our delivery. The average level of unique clones is around 20, with highest records reaching 50 plus. The affinities of the clones are also noticeable, ranging from 10-8 ~ 10-10 mol/L, even reaching 10-11 mol/L in some cases.
|Milestone||Service Spec||Delivery||Time (week)|
|1||Phage display panning and screening||Conduct up to 2 different panning strategies in parallel(2-3 subsequent round/panning)
ELISA test of phage clones for eight 96-well plates (~800 clones).
FACS Confirmatory Test.
Sequence all the positive phage monoclones to evaluate sequence diversity
|Monoclonal ELISA/FACS positive>50%
ELISA/FACS binding results
|3||Ab expression & purification||Select top 10-20 unique clones to full length antibody production||1mg purified Ab/ Sequence||3~4|
FACS binding and ELISA EC 50 (Up to 20 clones)
|Single point FACS binding test
ELISA EC50 for up to 20 clones
|5||Sequence Delivery (Up to 5 sequence)||Selected sequences (Up to 5) will be removed from library||Up to 5 sequences||/|
|Milestone||Service Spec||Delivery||Time (week)|
|1||Phage display panning||Conduct up to 2 different panning in parallel(2-3 subsequent round/panning)
ELISA/FACS on all phage clones
Sequence random 100 phage monoclones to evaluate sequence diversity
|Monoclonal ELISA/FACS positive>50% Sequence diversity>50%||3~4|
|2||FASEBA high throughput screening
|Construct FASEBA library by using the output phage;
ELISA/FACS test &SPR affinity ranking;
Sequence clones selected;
Affinity ranking report of affinity ranking;
|3||Full length Ab expression, purification & QC||Select top 10 unique clones to full length antibody production
QC by ELISA/FACS
|1mg purified Ab/ Sequence
*The positive rate in evaluation of Phage ELISA will be varied based on different targets, 50% shown in the flowchart is the average value and used as reference.
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