Module 9 2024

03/09/2024

Cell-Based Neutralisation Assays

Advantages

• Functional assay, which distinguishes antibodies with neutralising potential

• Provides information on cross-reactivity with other similar preparations

Disadvantages

•

Generally time-consuming

• Prone to non-specific matrix effects and interference with inhibitory molecules e.g., soluble receptors

UT7- Epo

•

Sensitivity can be an issue

250000

E1

E3

200000

• Optimisation and validation can be difficult

150000

cpm

100000

50000

0

10

100

1000

10000 100000

Reciprocal dilution

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Non-Cell-Based Neutralisation Assays

Competitive ligand binding assay (CLBA) can be used if relevant to the MoA of the therapeutic

MSD platform – assay principle

MSD platform - example

NAb loss of signal

binding ADA (non-NAb) signal

no ADA signal

NCB - adalimumab-treated patient

300

Sulfo-tag labelled therapeutic, incubated with sample (control Ab, matrix)

200

T0 T3 T6

100

ECL counts

Target immobilised (soluble Rec, ligand,…)

0

10

30

90

270 810 2430

reciprocal dilution

Adalimumab-treated patient serum samples (T0, T3, T6), incubated with biotin TNF  and sulfo-tag adalimumab.

Dotted blue line indicates binding of biotin TNF  and sulfo-tag adalimumab in absence of serum sample

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