On-line immunoaffinity solid-phase extraction capillary electrophoresis mass spectrometry using Fab´antibody fragments for the analysis of serum transthyretin

Talanta
Laura PontVictoria Sanz-Nebot

Abstract

This paper describes an on-line immunoaffinity solid-phase extraction capillary electrophoresis mass spectrometry (IA-SPE-CE-MS) method using an immunoaffinity sorbent with Fab' antibody fragments (Fab'-IA) for the analysis of serum transthyretin (TTR), a homotetrameric protein (Mr~56,000) involved in different types of amyloidosis. The IA sorbent was prepared by covalent attachment of Fab' fragments obtained from a polyclonal IgG antibody against TTR to succinimidyl silica particles. The Fab'-IA-SPE-CE-MS methodology was first established analyzing TTR standard solutions. Under optimized conditions, repeatability and reproducibility were acceptable, the method was linear between 1 and 25µgmL-1, limits of detection (LODs) were around 0.5µgmL-1 (50-fold lower than by CE-MS, ~25µgmL-1) and different TTR conformations were observed (folded and unfolded). The applicability of the developed method to screen for familial amyloidotic polyneuropathy type I (FAP-I), which is the most common hereditary systemic amyloidosis, was evaluated analyzing serum samples from healthy controls and FAP-I patients. For the analysis of sera, the most abundant proteins were precipitated with 5% (v/v) of phenol before Fab'-IA-SPE-CE-MS. The current meth...Continue Reading

Citations

Jan 25, 2020·Electrophoresis·Hana NevídalováZdeněk Glatz
Nov 3, 2020·Journal of Separation Science·Shannon L ThomasKatarína Maráková
Oct 27, 2020·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Chenchen LiuKoji Otsuka

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