Recombinant Antibody Production Services

Plasmid/vector construction, optional cell pool screening

We will synthesize the DNA sequence for the antibody's VH and VL variable regions and clone the gene into the expression vector. For transient expression, we will create an expression plasmid that's used to transfect the CHO cell line. For stable cell line expression, we will create a stable cell line (not included in the deliverables). Note, gene synthesis is calculated separately (US$ 0.2/bp)

Timeline: 1 week for transient expression, 2 weeks for stable cell line expression

recombinant expression

We will express the antibody in a fermentor (large scale) or cell culture (small scale), until we harvest enough antibody fragments for purification. We will validate with SDS-PAGE at this stage and ensure the purity measured by SDS-PAGE is >95%. Additional purification service can be ordered.

Timeline: 1 week for transient expression, 2-3 weeks for stable cell line expression

purification, characterization and Quality control

We will purify the antibody and characterize it for its affinity and titer. We will also ensure the endotoxin level is <1 EU/mg. Additional validation services can be ordered, as part of our CRO services, including Western blotting Service, ELISA service, IHC service, and Biocore SPR.

Timeline: 3-4 days

Delivery

We will prepare the antibody for final delivery.

Timeline: 1 week

Total timeline: 2 weeks for transient expression and 4-5 weeks for stable cell line expression. 1 week for delivery.

Antibody Formats

Format of antibody refers to the framework onto which the variable domains are engineered. The most commonly used antibody formats and their features and advantages are described below:

• Full-Length Antibodies (can switch isotype between IgG, IgM, IgA, and IgE): description
• Single-Chain Variable Fragments (scFv): description
  
• Fab (Fragment Antigen Binding) Fragments: description
  
• Single-Domain Antibodies (sdAbs): description
  
• Chimeric Antibodies: description
• Variable Domains Of Heavy-Chain Antibodies (VHH): description
  
• Bispecific and Brispecific Antibodies: description
  
• Antibody-Drug Conjugates (ADCs): description
  
• Fusion antibodies including Fc fusion proteins: description
  

Pricing: Compare Apples to Apples

The price per mg or per gram matters. However when comparing pricing one needs to make sure one is comparing apples to apples. Considers these parameters when comparing different offers for recombinant antibody production:

• Yield: total quantity of antibody produced, usually measured in miligrams or grams.
• Final pricing: pay attention to additional cost such as gene synthesis fees and shipping costs.
• Purity: it costs more to purify an antibody from 95% to 99% than it costs to purify from crudes to 95%.
• Endotoxin levels: low endotoxin levels is required for most in vitro and in vivo applications.
• Lead time: the latest technology on the market can go from sequence to mg scale of antibody in <2 weeks. Slower older technology tends to take 4+ weeks.
• As a quick reference, Boster's default recombinant antibody production service can produce 1 gram of antibody for $6000, gene synthesis and shipping included, in 4 weeks with 95% purity and <1 EU/mg endotoxin.

Turnaround Time

As mentioned above, the turnaround time or lead time is something to consider when selecting a partner. With the latest advancements in technology, the CHO cell recombinant antibody expression workflow has become a well established and widely available technology. Most competitive advantages are now business model based (instead of proprietary technology based). In other words, for a rather commodity service, the main way to cut down delivery time is to conduct gene synthesis in-house (a lot of smaller companies tend to outsource this, resulting in delays due to material transfer). Another common cause for delay is capacity limit. If turnaround time matters to you be sure to get confirmation of exact lead time, whether there is a queue to get started, when you receive a quote from vendors.

Endotoxin Levels And Purity

Endotoxin levels matter for recombinant antibody expression because endotoxins can trigger potent immune responses in mammals, potentially affecting the quality, stability, and safety of the produced antibodies. High endotoxin levels in recombinant antibody samples can lead to unwanted immune reactions and interfere with downstream applications, including in vitro cell based studies, in vivo studies and therapeutic use. It is essential to control and minimize endotoxin contamination to ensure the reliability and validity of antibody production and downstream applications.

Transient Vs. Stable Cell Line Expression

Transient expression is chosen for recombinant antibody expression when rapid production of small amounts of antibodies is required for screening or initial characterization. It is suitable for short-term studies and avoids the time-consuming process of establishing stable cell lines. At Boster Bio we can express 100+ antibodies for large scale candidate screening studies.

Stable cell lines are preferred for long-term production of large quantities of antibodies, ensuring consistency, scalability, and cost-effectiveness. They are suitable for therapeutic antibody production, high-throughput screening, and long-term research needs. The choice depends on the intended application, time frame, and scale of antibody production.

Antibody validation

Having the same company produce and validate antibodies offers numerous advantages and convenience. It ensures quality control throughout the entire process, minimizing variability and ensuring reliable results. Boster Bio is also a CRO that offers antibody characterization services such as ELISA to confirm specificity, Western blot to assess protein binding, flow cytometry for cell surface marker detection, and immunohistochemistry to verify tissue staining patterns. Validations also encompass functional assays, such as neutralization assays for therapeutic antibodies. A one-stop solution simplifies communication, reduces logistical challenges, and provides a comprehensive and streamlined approach to antibody development and validation.

Antibody Engineering: Humanization and Affinity Maturation

Boster Bio is able to provide antibody engineering services as well. Seamless integration of production and engineering streamlines the process, ensuring rapid and efficient modifications. Customized humanization and affinity maturation can be tailored to the specific needs, optimizing antibody properties for therapeutic applications or research. Close collaboration between production and engineering teams enhances communication and accelerates project timelines. Overall, the integrated approach simplifies project management, minimizes coordination efforts, and delivers high-quality engineered antibodies tailored to precise requirements.