What is the typical timeline for a VHH antibody discovery project?

A typical project will take 4-5 months. The timeline varies based on project complexity and specific requirements. Generally, the process includes antigen preparation, animal immunization, library generation, candidate screening and validation, and functional assays.

Can Biointron work with specific antigens provided by clients?

Yes, Biointron can produce antigens, or work with antigens provided by clients.

What validation methods are used for VHH antibodies?

Biointron employs multiple assays for validation, including ELISA, SPR, and FACS, ensuring the specificity of the discovered antibodies.

Can Biointron obtain VHH antibodies with a wide range of affinities?

Yes, utilizing phage display technology and tailored biopanning methods, Biointron generates VHH hits with affinities ranging from 10⁻⁸ to 10⁻¹¹ M.

What are the deliverables at the end of a VHH antibody discovery project?

Clients receive detailed reports, including discovery process, sequence information, binding affinity data, functional assay results, and purified VHH antibody samples for further testing.

Can Biointron support VHH developability after discovery?

Yes, Biointron offers developability assays to support the development of VHH antibodies for both therapeutic and diagnostic applications.

How does Biointron handle intellectual property rights for discovered VHH antibodies?

Yes, all intellectual property rights for antibodies discovered under client-specific projects belong to the client.

How can I initiate a VHH antibody discovery project with Biointron?

To start, contact Biointron through the website’s inquiry form or email. A meeting will be arranged with a Biointron team member to discuss project requirements and provide a tailored proposal.

VHH Antibody Discovery Services

Accelerate your VHH antibody discovery with Biointron’s 4-5 month timeline.

At Biointron, each program starts with a dedicated alpaca from our 300+ herd to build a deep, target-focused immune response. We construct high-diversity VHH libraries (≥10⁸) and run tailored phage display campaigns to maximize hit rate and epitope coverage. Selected VHHs are expressed and validated by ELISA/SPR/FACS (plus internalization or ADCC when format-appropriate), routinely yielding KD values across ~10⁻⁶–10⁻¹¹ M (assay- and target-dependent). Use your own antigen or have us produce it. Deliverables include a full report, sequence files, affinity/assay data, prioritized hit/lead tables, and purified VHH samples—plus next-step recommendations. All antibodies generated under your project belong to you.

Our antibody discovery services also include:

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Transform your research with Biointron's VHH Antibody Discovery Services. From our exclusive alpaca breeding farm to our advanced phage display technology, Biointron delivers high-diversity, single-domain VHH antibodies tailored to your project needs. With 100+ successful projects annually and a dedicated team of experts, we provide a consistent, reliable solution for therapeutic, diagnostic, and environmental applications.

Advantages of our VHH Antibody Discovery Platform

Self-owned Alpaca Breeding Farm

300+ alpacas available
One alpaca just for one project

100+ projects delivered/year

Various immunization/panning/screening methods
Multiple assays for validation (ELISA, SPR, FACS, Internalization, ADCC, etc.)

High Diversity and Large Capacity

Maximizing success with high diversity and large capacity
Library size at least 10⁸ - 10⁹

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Scale, Speed, and Quality—All Under One Roof

Transform your research with Biointron's VHH Antibody Discovery Services. From our exclusive alpaca breeding farm to our advanced phage display technology, Biointron delivers high-diversity, single-domain VHH antibodies tailored to your project needs. With 100+ successful projects annually and a dedicated team of experts, our integrated workflow takes you from antigen to qualified VHH leads with robust QC and proactive communication at every milestone.

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End-to-End VHH Programs, Built for You

We combine dedicated-animal immunization, immune library construction (≥10⁸), and target-specific phage display with orthogonal screening to deliver high-confidence binders and ranked leads. Seamless downstream support—humanization, affinity maturation, developability, and small- to large-scale production—helps you advance rapidly into functional studies, formatting, and preclinical evaluation.

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Biointron Alpaca VHH Affinity Distribution

0%AF-5
4%AF-6
16%AF-7
33%AF-8
32%AF-9
13%AF-10
2%AF-11
0%AF-12
0%AF-13

Data source: 615 VHH antibodies (as part of our developed VHHs)

Calculation example:

Total number of VHH of affinity 10^-9 M from different target

/

Total number of VHH of from different target (615 unique sequence)

=

32%

With the Biointron VHH discovery platform, VHH hits with affinity ranging from 10^-8 to 10^-11 M have been generated. It should be noted that the presented data represents only a portion of projects from one client and may have sampling bias. The distribution of hits with different affinity may not reflect the variance between different targets. The data is from SPR affinity measurement.

VHH Antibody Discovery Workflow

Antigen expression or provided by client

Animal immunization

PBMC isolation

mRNA Extraction and Reverse Transcription

Library generation

Library screening and Tailored biopanning ( solid phase panning & liquid phase panning)

Positive Clone Sequencing and Sequence Analysis

VHH expression

VHH Validation

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Applications of VHH Antibodies

VHH antibodies have been utilized in a wide variety of settings, such as clinical therapeutics and immunodiagnostics, as well as environmental monitoring, due to their differentiated characteristics. For example, Caplacizumab is an antitumor therapeutics for blood disorders, while CAR-T therapy benefits from VHH-based CAR constructs to enhance treatment precision.

VHH Antibody Discovery Service Details

Our phased approach provides structured, reliable antibody discovery:

Service Step	Service Description	Timeline	Deliverables
Phase Ⅰ: Antigen preparation / Antigen validation	Antigen can be prepared by BioIntron or provided by our client	2 weeks for preparation / 2-3 days for validation	1mg purified protein antigen If antigen provided by client, it should be at least 4mg with SDS-PAGE >95%, endotoxin level <1EU/mg
Phase Ⅱ: Alpaca immunization and PBMC isolation /preservation	5ml pre-immune bleed the ELISA titer after 2nd, 3rd and 4th immunization will be monitored and the report will send to client the 50ml PMBC after 2nd, 3rd and 4th immunization will be extracted and stored in Trizol	8-10 weeks	Alpaca's death caused by antigen, there will be setup fee about $5,000 Decision gate: based on the ELISA titer on the 2nd, 3rd and 4th immunization. Our client can decide to use which cell for library generation
Phase Ⅲ: VHH Phage Library Generation	RNA extraction and cDNA preparation VHH amplification Electro competent E. coli Library Diversity and capacity measurement clone NO counting and sequencing to measure the capacity of the library	3-4 weeks	VHH capacity reach to 10⁸ - 10⁹
Milestone Ⅳ: Library Biopanning and Screening	at least 3 round panning (liquid or solid phase panning) ELISA verification, identification and evaluation of positive clones screening positive clone sequencing and sequencing result analysis	3-4 weeks	Guarantee to provide at least 20 unique sequence
VHH-Fc Antibody Production	Gene synthesis, subcloning, plasmid preparation transient expression and purification QC analysis	2 weeks	Purified antibody (SDS-PAGE >95%, endotoxin level <1EU/mg)
Affinity Ranking (optional)	Affinity measured by Biacore 8K	0.5 weeks

Phase Ⅰ: Antigen preparation / Antigen validation
Timeline

2 weeks for preparation / 2-3 days for validation

Description
- Antigen can be prepared by BioIntron or provided by our client
Deliverables
- 1mg purified protein antigen
- If antigen provided by client, it should be at least 4mg with SDS-PAGE >95%, endotoxin level <1EU/mg
Phase Ⅱ: Alpaca immunization and PBMC isolation /preservation
Timeline

8-10 weeks

Description
- 5ml pre-immune bleed
- the ELISA titer after 2nd, 3rd and 4th immunization will be monitored and the report will send to client
- the 50ml PMBC after 2nd, 3rd and 4th immunization will be extracted and stored in Trizol
Deliverables
- Alpaca's death caused by antigen, there will be setup fee about $5,000
- Decision gate: based on the ELISA titer on the 2nd, 3rd and 4th immunization. Our client can decide to use which cell for library generation
Phase Ⅲ: VHH Phage Library Generation
Timeline

3-4 weeks

Description
- RNA extraction and cDNA preparation
- VHH amplification
- Electro competent E. coli
- Library Diversity and capacity measurement
- clone NO counting and sequencing to measure the capacity of the library
Deliverables
- VHH capacity reach to 10⁸ - 10⁹
Milestone Ⅳ: Library Biopanning and Screening
Timeline

3-4 weeks

Description
- at least 3 round panning (liquid or solid phase panning)
- ELISA verification, identification and evaluation of positive clones screening
- positive clone sequencing and sequencing result analysis
Deliverables
- Guarantee to provide at least 20 unique sequence
VHH-Fc Antibody Production
Timeline

2 weeks

Description
- Gene synthesis, subcloning, plasmid preparation
- transient expression and purification
- QC analysis
Deliverables
- Guarantee to provide at least 20 unique sequence
- Purified antibody (SDS-PAGE >95%, endotoxin level <1EU/mg)
Affinity Ranking (optional)

Timeline

0.5 weeks

Description

Affinity measured by Biacore 8K

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Case Study

Biointron’s proven process has achieved substantial outcomes.

Case 1: Anti-X Antigen

After panning and screening, 53 unique sequences were obtained & expressed in the HTP CHO expression system.

Validation of VHHs Binding to X (by ELISA)

49 of 53 VHH Antibodies bound to X (ELISA)

Validation of VHHs affinity with X (by SPR)

5No Binding
9E-7
21E-8
16E-9
2E-10

Affinity range 5.36x10^-7 M to 7.66x10^-10 M.

48 of 53 VHH Antibodies bound to X (SPR)

Validation of Cell-based VHHs Binding to X (FACS)

43 of 53 VHH Antibodies Bound against CHO-Kl/X Cell

VHHs Agonist Activity Validation

31 of 53 antibodies showed significant agonist activity

VHHs Validation (Blocking assay)

18 of 53 antibodies showed significant blocking activity.

Case 2: VHH Antibody Discovery (Target: Claudin 18.2)

Alpacas were immunized with Claudin 18.2-encoding DNAs, followed by immunizations with Claudin 18.2-overexpressing cells. Alpaca serum taken after immunization manifested specific binding to Claudin 18.2-overexpressing cells (but not to parental ones).