One-Stop Solutions for Antibody Drug Discovery

GenScript Solutions

Antibody Sequencing (Sanger & NGS) are reliable choices to speed up therapeutic antibody lead discovery and obtain accurate antibody sequences.

Validated antigens (DNAs/RNAs , peptides , proteins , and cell lines/pools ) are leveraged to generate therapeutic antibody candidates.

GenExact™ ssDNA is a non-viral template for CRISPR-mediated KI that may be leveraged to integrate recombinant V(D)J regions to generate transgenic models for antibody production.

MonoRab™-B Cell Cloning Platform is a novel discovery platform that delivers rabbit monoclonal antibodies with high specificity & affinity for your lead generation.

Beacon-Single B Cell Sorting Platform enables achieving the most diversified antibody repertoire to capture all opportunities with a high success rate.

Solution Highlight 1
Multiple immunization strategies

Solution Highlight 2
Developing therapeutics at pandemic speed and with high affinity and diversity

Case Study

• Case Study 1: MonoRab™-B Cell Cloning Platform

  Workflow of MonoRab™-B Cell Cloning Platform

  

  Antigen: Human 4-1BB ECD,  24-186 AA

  Results:

  107 protein positive clones

  43 Human and Cyno 4-1BB FACS double positive clones and 9 Human, Cyno and Mouse 4-1BB triple FACS positive clones

  107 clones for NGS sequencing

  43 clones for antibody transfection and validation and 33 clones validated by ELISA and FACS

  High success rate obtaining Abs with Human, Cyno and Mouse cross-reactivity.

  
• Case Study 2: Beacon-Single B Cell Sorting Platform

  Workflow of Beacon-Single B Cell Sorting Platform

  

  Antigen: Human 4-1BB ECD,  24-186 AA

  Results:

  1191 positive wells from bead assay

  267 positive wells from cell-based assay

  50 clones for NGS sequencing and 31 clones with successful VH and VL pairing

  Single B cell level functional assay and Higher positive rate (>500 hits/chip)

• Case Study 3: Antibody Sequencing (Sanger & NGS)

  107 clones from previous B cell cloning platform were sequenced by NGS.

  Results:

  Out of 107 clones, 5 clones share same V-region with other clones.

  Antibody sequence analysis indicates antibody diversity and identifies candidate.

  

  

  

GenScript Solutions

CellPower™ assay cell line is an appealing tool for cell-based binding assays to evaluate lead efficacy in a cellular environment with the particular advantages of predictability, automation, multiplexing, and miniaturization, supporting the drug discovery process.

Ab genes to vectors is a seamless delivery of candidate genes in your selected expression vector.

CRISPR mediated cell engineering could be performed as disease-specific knockout, knock-in, and inducible expression to evaluate lead efficiency or generate disease models to facilitate discovery.

Precision mutant library provides superior variant coverage in your highly diverse mutant pools to reduce the screening burden, powered by GenTitan Oligo Pool ^New.

TurboCHO™ high throughput/express is the most cost & time-effective (Only 2 weeks) option for small/middle-scale recombinant antibodies and a better solution for therapeutic target screening and optimization.

Solution Highlight 1
Quick filter-Gene to Plasmid

Solution Highlight 2
High throughput, rapid screening of affinity matured antibodies-Precise Mutation Libraries

Solution Highlight 3
TurboCHO™ High throughput and express antibody expression platforms

Whether it is a hybridoma platform or B cell cloning technology, the molecular sequences of the final candidates need to be prepared, expressed, and the resulting recombinant antibodies are further screened and verified. How to construct expression plasmids cost-effectively at high throughput is a problem that must be solved in the research process. GenScript Gene to plasmid service helps researchers to express and screen antibodies more quickly and economically.

To engineer the CDR regions of light and heavy chains in antibody candidates, GenScript provides a complete mutant library for affinity maturation. In a previous case study, the 6 CDR regions on the heavy and light chains contained 63 mutation sites. Through precise mutations for saturation scanning, each wild-type amino acid codon was mutated to the other 19 amino acid codons. Several high-affinity mutants were obtained, some of which had improved affinity, with up to 4 orders of increased magnitude. Using different types of precise mutation libraries at different stages can efficiently improve antibody affinity maturation.

Mammalian expression systems are crucial to ensure proper post-translational modifications and protein folding. CHO cells are a commonly used host system for expressing recombinant antibodies. In the process of candidate optimization, multiple antibodies need to be expressed simultaneously for validation. By relying on GenScript’s TurboCHO™ High Throughput platform, antibody production is significantly expedited. In 2 weeks, this platform delivers a range of purified antibody products (e.g., IgGs, Fab, scFv, VHH, bi-specific antibodies, and more) in quantities from µg to mg, and the Express platform could deliver quantities from 5 mg to 500 mg in 3 weeks.

General Workflow of TurboCHO™ High throughput Services

Case Study

• Case Study 1: High Throughput and Express Performance

  TurboCHO™ system can deliver a range of purified antibody products. M: Protein Marker; R: Reducing Condition; NR: Non-reducing Condition.

  

  

  
• Case Study 2: Bispecific antibody pairing can be validated by additional Mass Spectrometry QC

  Bispecific antibodies (like Knob-into-hole, KIH) play an important role in drug development, as this pioneering format enables binding to 2 different antigens (e.g. cell surface proteins or ligands), thus directing therapeutic agents to the intended target. GenScript's proprietary backbone along with well established multi-step purification platform enables us to generate bispecific antibodies with high purity.

  

  

  

GenScript Solutions

Anti-idiotype antibody discovery supports the development of reagents for drug development assays (PK, ADA, diagnostic and biomarker discovery and evaluation). 

TurboCHO™ High Performance supports the high yield needs of functional and therapeutic studies during the late stages of the antibody drug discovery workflow, with yields of up to 2.5 g/L, for best consistency and minimal surprises.

Custom ELISA Kits - PK/ADA/HCP specifically developed kits that enable the reliable detection and quantitation of your unique antigen.

Case Study

• Case Study 1:MonoRab™ for Anti-idiotypic antibody development

  One round of B cell cloning to generate Anti-ID blocking mAb

  • 154 clones: antigen positive, total HIgG negative
  • 81 clones: validation of binding specificity
  • 10 clones: blocking rate >80%

  

  
• Case Study 2: TurboCHO™ High Performance antibody expression

  TurboCHO™ HP Service Timeline

  

  High Expression Titers for 6 mAbs with Average Titer Reaching 3.1 g/L.

  

  Rabbit mAb production tested in the TruboCHO™ HP platform reached 2.1 g/L average yield, and >97% purity for all 3 molecules.

  mAbs	pI	MW/kD	EC	Purity by SEC-HPLC(%)	Purity by PAGE(%)	Yield (mg/L)
  mAb-1	7.59	140.5	1.481	98	90	2374.82
  mAb-2	7.15	143.2	1.430	97	90	1971.26
  mAb-3	8.06	142.1	1.429	98	95	1951.61

  

  TurboCHO™ system could reduce the levels of N-linked mannose-5 glycan (Man5), <5%, which helps to enhance antibody-dependent cell-mediated cytotoxicity (ADCC), compared to the previous CHO system.

  

  
• Case Study 3: ELISA Kit Development, including ADA kits, HCP kits, PK kits and conventional kits

  PK kits developed by GenScript meet or exceed FDA requirements

  

  Parameter	GenScript
  Linear range	0.5 ng/ml~24ng/ml
  ULOQ	24 ng/ml
  LLOQ	0.5 ng/ml
  Selectivity	80%
  Precision	Intra assay CV<10%
  	Inter assay CV<10%
  Recovery/ Interference Studies	80-120%
  Specificity/Cross-Reactivity	Not Done
  Matrix	Human Serum

  HCP ELISA Kit - CHO developed by GenScript meets or exceeds FDA requirements

  

  Parameters	GenScript
  Linear range	1-100 ng/mL
  Sensitivity	0.608 ng/mL
  Precision – Intra assay Variation	≤5%
  Precision - Inter-assay variance	≤10%
  Accuracy-Recovery	85-115%
  Recovery/ Interference Studies	80-120%
  Specificity	Drug
  Matrix	Injection salt solution