AOAC SPIFAN Stakeholder Panel Meeting Book (September 7, 2019)

Measuring lactoferrin content

X Some important questions:

X What does the method measure? X Intact/Native protein X Denatured protein X Fragments of the protein X Post-translational modifications (PTMs) X Glycosylation variable and prevalent

X What is our standard? X What is our sample? X How do we correlate the two?

Karav S., German J.B., Rouquié C., Le Parc A., Barile D. Studying Lactoferrin N-Glycosylation. Int. J. Mol. Sci. 2017;18:870. doi: 10.3390/ijms18040870

Available methods/techniques

Optical biosensor (Indyk et al, 2005)

X

LC-MS/MS (Ellingson et al, 2019/Zhang et al, 2014)

X

X RP-HPLC-UV (Drackova et al, 2009; Tsakali et al, 2014; Billakanti et al, 2019; Palmano et al, 2002, Pochet et al, 2018; Tsakali et al, 2014, 2019)

Heparin affinity (Zhang et al, 2017)

X

Ion Exchange chromatography (Billakanti et al, 2019)

X

CE-UV (Li et al, 2012; Mao et al 2017)

X

SDS-PAGE (Ronayne de Ferrer et al, 2000)

X

SDS-CGE (Feng et al, 2017)

X

X ELISA (Davis & South, 2015; Hiss et al, 2008; Wang et al, 2018)

Immunosensor (Campanella et al, 2009)

X

X FP aptasensor - Silver-Enhanced Fluorescence Polarization (Chen et al, 2017)

Cleanup/purification:

Affinity chromatography

X

Size exclusion chromatography

X