Interference caused by endogenous components of the sample

Also natural existing proteins of the specimens can cause interference.
Well-known interfering substances in human sera are: albumins, complement, lysozyme and fibrinogen. Endogenous proteins can bind assay antibodies or target analyte and so mask them. Substances can be demasked by using LowCross-Buffer® as assay buffer.

cross-reactivity with capture and detection antibody — Cross-reactivity with capture and detection antibody. In practice it is an uncommon phenomenon, but possible with antibodies of low specificity. Such interference can be caused by antibodies which are targeted on conserved amino acid sequences of a protein. These conserved amino acid sequences can also be found in other (therefore interfering) proteins.

binding analyte specimen protein — Masking of the analyte by a specimen protein. The capture antibody can not bind the epitope. Binding of analyte is not possible or bad (in case of sterical hindrance). The results are false negative signals.

Supplementary Methods, Troubleshooting, Regulatory Requirements
Editors: Arnold Maria Raem, Peter Rauch

This methods book provides a well-structured and accessible introduction to the fundamental principles of immunological methods.
Scientists in the life sciences and medicine,…

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Interference caused by endogenous components of the sample

Immunoassays – 1st Edition

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