Antibody characterization services: which measurements actually decide a candidate, what a human antibody or humanised candidate owes on top of them, what to establish before you buy antibody material or antibody buy decisions are made at scale, and the order to buy them in

The order to buy measurements in

1. Specificity before affinity. An antibody that binds tightly to the wrong thing is worse than one that binds moderately to the right thing. Screen against a counter-panel of close relatives and the species you will work in before spending on kinetics, since this eliminates candidates fastest and most cheaply.
2. Affinity and kinetics with the method stated. Surface plasmon resonance and biolayer interferometry give association and dissociation rates as well as the equilibrium constant, and the off rate is frequently what matters functionally. Ask which method, which orientation and which regeneration conditions, because all three move the numbers.
3. Epitope binning when you have several. Grouping candidates by whether they compete tells you how many distinct epitopes you actually have and which pairs could work together in a sandwich assay or a bispecific. It is worth doing once the panel is down to a manageable number.
4. Function in the assay that matters. Binding does not predict blocking, agonism, internalisation or staining behaviour. A functional assay in the intended format is the measurement that decides the candidate, and it should be planned even if it is run in house.
5. Developability, before you commit. Thermal stability, aggregation propensity, expression level and behaviour at concentration decide whether a candidate can be made and stored. These are cheap to screen and expensive to discover late.

What a report should state

The method, the instrument, the orientation, the buffer, the temperature and the analysis model, alongside the numbers. Affinity values are method dependent and are not comparable between reports that do not state these things.

Ask for the raw sensorgrams or traces. A fitted constant without the underlying curve cannot be assessed, and poor fits are common.

Reference standards and reproducibility

Include a well-behaved reference antibody in every run so that results are comparable across days and across providers. It is the cheapest way to detect that something in the assay has moved.

Retain aliquots of every candidate measured. Re-measuring later without the original material tells you nothing about whether the antibody or the assay changed.

Common questions

What should I measure first on an antibody panel?

Specificity against close relatives and the working species. It eliminates candidates fastest and most cheaply, and a tight binder to the wrong target is worse than a moderate binder to the right one.

Are affinity values comparable between providers?

Only when the method, instrument, orientation, buffer, temperature and analysis model are the same. Ask for all of those alongside the number, and for the raw traces.

What is epitope binning for?

Grouping candidates by competition, which tells you how many distinct epitopes you have and which pairs can work together in a sandwich assay or a bispecific format.

Why screen developability early?

Because thermal stability, aggregation propensity and expression level decide whether a candidate can be made and stored at all. They are cheap to screen and very expensive to discover after lead selection.

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