Why use Single-chain antibodies?

Immunoglobulins from camelids are sub-classified as IgG1, IgG2 and IgG3.
IgG2 and IgG3 are single-chain Abs, and they both bind the antigen through a unique domain, named VHH (or nanobody®).

They differ by the size of their hinge region which connects the VHH domain to the constant IgG domain (CH2 + CH3).
It results in a size of 90 to 100 kd, instead of ~ 150 kDa for a classical IgG1.

Simpler structure

IgG2 and IgG3 are both devoid of light chains or CH1 domains. Hence their size (~90 kDa) is much smaller than the classical IgG1 size (~150 kDa).

As some disulfide bonds (connecting the heavy and light chains in conventional Abs) are missing. 

HcAbs offer a much easier manipulation and a higher stability. HcAbs are e.g. reported to bind antigens at higher temperatures and under more stringent denaturing conditions compared to classical IgGs.

Selectively working with the IgG2/3 serum fraction of an immunized llama is made possible by the availability of anti-IgG2/3 secondary antibodies, which show no cross-reaction toward IgG1.

VHH fragments and their benefits

VHHs have many advantages over traditional immunoglobulin fragments such as scFvs or Fabs.

 

Compared to monoclonal antibody fragments, VHHs are made of one polypeptide chain only. Therefore, they can easily be produced in lower organisms (E. coli, yeast). They also retain full activity compared to the HcAb from which they originate.

On the opposite, Fabs, scFvs and other mAb fragments are made of two polypeptide chains, and their folding without the supporting scaffold of the original mAb may lead to specificity or sensitivity issues. Producing dimeric fragments is also more challenging.

Benefits

Better targeting

Due to their small sizes (~15kDa), VHH antibody fragments can bind epitopes that are hidden, so the targeting precision is higher compared to normal antibodies.

Benefits

Easier production

Because they are composed of a unique polypeptide chain, VHH are relatively easy to produce in lower eukaryotes up to very high amounts and purities.

Benefits

Better diffusion

VHH have a high tissue penetration and are cleared from circulation rapidly. Some VHH can even cross the blood-brain barrier.

Benefits

Higher stability

VHH are stable under extreme pH, temperature and against proteases; hence, they keep their native folding and epitope binding capacity under very diverse experimental conditions.

Our camelid antibody production services

We offer llama and alpaca immunization and the production of custom VHH antibody fragment.

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Anti-llama secondary antibodies

We provide labeled (biotin, R-PE or custom labeled) and unlabeled secondary antibodies to target llama IgG1 or IgG2/3 immunoglobulins with high precision and sensitivity.

We also offer a unique, top-quality secondary antibody targeting VHH’s.  This rabbit polyclonal is broad VHH-specific, as it binds to and allows the detection of VHHs from different germline sequence families.

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