NGS
Antibodies
IVDR Solutions
Twist Bioscience HQ
681 Gateway Blvd
South San Francisco, CA 94080
Technical Documents
How it works
Walk through our standardized workflow
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Transient expression
Robust, reproducible protein production. Parallel expression across 96 deep well plates.
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Purification
Protein A or His-tag batch purification of anitbodies. Purity, size, concentration, and yield data for every antibody.
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QC & Characterization
Optional endotoxin testing, binding, developability and custom assays available.
Characterization specsCheck performance data
Review yields and QC outcomes from recent production runs. Compare HEK293 against CHO, and see the quality standards every order meets.
Figure description
A diverse panel of single-chain VHHs (Fc-fusion and 6xHis-tagged) and full-length IgGs from multiple discovery campaigns were expressed in Expi293™ cells to demonstrate expression variability. Antibodies were produced in 1.2 mL reactions, purified, and quantified by A280, with triplicate transfections ensuring reliable, plate-to-plate performance comparisons.
Figure description
The same antibody panel was expressed in ExpiCHO™ cells (1.2 mL scale) to demonstrate expression variability across constructs and purification types. Antibodies were purified and quantified by A280, with triplicate transfections enabling reliable plate-to-plate comparisons.
Quality antibodies for discovery
To ensure that antibodies are ready for experimentation “out of the box”, quality control (QC) of purified antibodies includes A280 quantitation, size (kDa) and percent purity analysis with a denatured capillary electrophoresis sodium dodecyl sulfate (CE-SDS) assay. After passing QC, the antibody is prepared for shipping unless buffer exchange or other downstream processing is requested. Shown below are typical QC results generated using Twist's High Throughput Antibody Production workflow. The clean peaks in (A) and bands in (B) show that species within the antibody stock are all of similar size and charge. The peak at 67 kD represents the heavychain segment of the antibody, while the peak at 29 kD represents the light-chain segments.
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Expression system specs
How to order
Place your order in three steps
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Download & complete template
Download the template to get started. Include sequences, vectors, kozak, leader, and purification type.
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Upload completed template to your order
Start your antibody order and upload the completed template or paste in names and sequences.
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Select your order settings
Choose your scale, elution buffer, buffer exchange, QC, and delivery format.
Ready to get started?
Start your design now or speak with an expert to scope your project.
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Antibody Characterization
Characterization data in as few as 15 business days
Antibody characterization assays
Our antibodies and services are just one part of the workflow
Pair our high-throughput antibodies and services with a full range of versatile Twist solutions.
Antibody Discovery Services
Partner with our team of experts to navigate the entire discovery funnel from target to lead.
Oligo Pools
High-diversity, silicon-synthesized oligo pools designed for high-throughput screening and complex library construction.
Libraries
Explore vast sequence space with high-diversity libraries where every variant is base-by-base synthesized and NGS-verified to match your design.
Genes
Explore vast sequence space with high-diversity libraries where every variant is base-by-base synthesized and NGS-verified to match your design.
FAQ
General
Does Twist supply prepped DNA along with purified or supernatant antibodies?
Twist supplies clonal gene glycerol stocks from the clonal genes synthesized for antibody production. During antibody production, the DNA is consumed for transfection; however, glycerol stocks are reserved and delivered along with the antibodies post-production. Please see our protocol for preparing fresh DNA from glycerol stocks.
What does a leader (signal peptide) sequence do and what leader sequence do we use in our pTwist vectors?
A leader or secretory signal sequence acts as a peptide signal bound by cellular machinery early in the translation process. This allows for the correct localization of the translation complex to the endoplasmic reticulum where the antibody chains can enter into the secretory pathway for protein assembly and secretion from the cell. The leader sequence is cleaved during the protein assembly process and is not a part of the final protein. Twist recommends the following leader sequence for both heavy and light chains when using our pTwist IgG expression vectors as these sequences have been tested and validated with our pTwist vectors in both Expi293™ and ExpiCHO™ cell lines:
VH: MRAWIFFLLCLAGRALA
VL: MRAWIFFLLCLAGRALA
What is a Kozak sequence, and what is the Kozak sequence used when I select "Twist sequence" during antibody design
The Kozak sequence directs the pre-initiation complex (PIC) and ribosome to the translation initiation site (start codon) and mediates ribosome assembly ensuring the correct protein sequence is translated.
The consensus Kozak sequence is generally considered as GCCGCCACCATGG, where ATG is the start codon (typically the start of the signal sequence). Twist recommends the version GCCGCCACC upstream of the start codon ATG.
I am receiving my samples in media as Supernatants, how do I purify my antibodies?
You can use a Protein A or G solid phase to purify, which binds to the FC region of the antibodies, depending on species and isotype of IgG. Alternatively, you can use immobilized antigen (full length or short peptide antigen) to purify the antibodies. The method will depend on the downstream application and specificity of the antibody.
What strain of cells was used to produce these antibodies?
Twist uses the industry standard cell lines, Thermo Fisher Scientific Expi293™ and Thermo Fisher Scientific ExpiCHO™
Can I have an antibody made with a custom Fc region such as IgG3 Fc, mutated IgG 1 Fc or other species such as mouse?
Customers may select the pTwist CMV BG WPRE Neo expression vector when they would like to use their own Fc region sequence (IgG3, Mouse IgG, or modified Fc) or have another type of Fc-fusion antibody to express.
Please remember to include a stop codon at the end of your coding sequence. If you have any design questions, please email igg@twistbioscience.com for help with your custom request.
Why are the C-terminal lysine residues missing from our IgG constructs?
Research has shown that C-terminal lysine residues of the heavy chain (present on all IgG isotypes) are cleaved off once the IgG is in circulation. Studies have also shown that there is no functional impact of the lysine once it's cleaved and its presence is actually inhibitory to allowing the antibody to reach full cytotoxicity potential: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4622059/
Why is the 10th positon cystein in our VHH-Fc fusion vectors mutated to a serine?
This cystein in a full IgG forms a disulfide bond with the LC constant region which is not present in the VHH-Fc fusion construct.
How should I upload my antibody sequences?
Please ensure you have the following sequence format for uploading variable regions:
- Removed signal leader peptide (if using pTwist vectors + Twist recommended signal leader peptide)
- Variable regions should not end with any CH1 starting sequence
- Sequences are divisible by three if your ORF is in line with your insertion site
What is our pricing breakdown for custom antibodies?
Delivery Format
What formulation and buffer exchange options are available for antibody shipments?
Antibodies are typically supplied in standard commercial Phosphate-Buffered Saline (PBS). Depending on your application parameters, you can specify alternative pH preparations—such as PBS at pH 7.4 or custom configurations like PBS at pH 6.0—directly through the portal configuration window.
What are the expected delivery volumes and yields for the 1 mL and 8 mL production scales?
Final metrics vary based on individual antibody sequence expression characteristics. Standard yields break down across the following parameters:
- 1 mL Scale: Delivers a final elution volume of approximately 220 µL, with target yields ranging from 15 µg to 300 µg.
- 8 mL Scale: Delivers a final elution volume of approximately 750 µL, with target yields ranging from 15 µg to 3,000 µg.
What physical layout options and vial formats can be selected for final delivery?
To ensure seamless compatibility with downstream laboratory automation systems, microfluidics arrays, and fluid handlers, products can be specified in several array configurations:
- 96-deep well plates featuring fully customizable grid array mapping.
- 1.4 mL matrix-compatible micronic tube racks.
- Individual specialized tubes available in 2 mL, 4 mL, or 10 mL capacities (10 mL tubes are optimized for large volume supernatant shipments).
Resources and guidance for sequence design
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abYsis provides an integrated, comprehensive, and annotated sequence assessment for one or multiple
antibodies. abYsis provides a thorough regional breakdown of antibody sequences to quickly identify where
your
variable region starts and ends. A tabular breakdown of the leader, FWK, and CDR sequences as well as the
tail, or constant region sequence is also provided.
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It can be helpful to apply the annotations provided by abYsis to your sequence using bioinformatic programs to help visualize and isolate your variable region. Below is an example of a sequence imported to the Geneious Prime software platform with abYsis annotations applied.
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One additional option that may provide comprehensive breakdown of leader, variable, and constant regions is ANARCI. The resulting analysis provides the exact variable region sequence highlighted in red, which can then be uploaded to the Antibodies application on our eCommerce website. ANARCI also provides detail on the light chain domain type and the sequence species.
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Please keep in mind that variable region sequences should never start with a methionine (M). If you are uploading sequences that start with the amino acid M, or nucleotides ATG, and are planning to use pTwist vectors, please contact our support for assistance.
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Reformatting variable domains from one species to another can be complicated given differences between disulfide bonding and other features. By default, the Twist eCommerce platform will not correct any of these issues. We typically rely on you to account and inspect for these types of reformatting complications. If you need assistance, please contact our support team.
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Collaborate with a trusted scientific partner to guide your project specs. From bespoke data configurations to workflow design, our experts provide the proactive support you need to reduce friction at every step.
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For Research Use Only.
Not for Diagnostic Procedures.
*Terms & Conditions: Turnaround time starts at 10-15 business days for 1mL antibody expressed in HEK293 and 13 - 18 business days for 1mL antibody expressed in CHO. 8 mL expression volumes take an additional business day.
Pricing is quoted in U.S. dollars and all turnaround times are based on U.S.-based operations, business days, and service availability. Pricing and turnaround estimates are not guaranteed for international orders and may vary based on location, currency conversion, customs, taxes, duties, local regulations, shipping conditions, and other factors outside our control. Customers outside the United States are responsible for confirming applicable pricing, fees, and timelines before purchase.
