Ficolin-2 reveals different analytical and biological properties dependent on different sample handling procedures

Hein, Estrid; Bay, Jakob Thaning; Munthe-Fog, Lea; Garred, Peter

doi:10.1016/j.molimm.2013.05.233

Cited by 25 publications

(30 citation statements)

References 32 publications

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“…Our study confirmed a recent finding by a European group, which was published while we were preparing our report (26). However, we found that the effect of tube type on the serum level is dependent on the assay type, since we observed that equivalent levels of ficolin-2 were demonstrated by Western blotting for ficolin-2 from sera collected in four different tube types (Fig.…”

Section: Discussionsupporting

confidence: 92%

Blood Collection Tubes Influence Serum Ficolin-1 and Ficolin-2 Levels

Brady¹,

Spencer²,

Falsey³

et al. 2013

Clin. Vaccine Immunol.

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The ficolins are members of a recently discovered family of host innate opsonins that can activate the lectin pathway of complement. The ficolins bind many ligands, although they are typically described as binding acetylated sugars. Ficolin-1 (M-ficolin) and ficolin-2 (L-ficolin) are known to bind Streptococcus pneumoniae serotypes 19C and 11A, respectively. While studying the binding of ficolins to pneumococci, we found variations in ficolin-2 binding among serum samples collected in different types of blood collection tubes. Plastic tubes, which contain a silica clot activator, yielded sera with reduced ficolin-2 binding and apparent ficolin-2 levels. We found that the silica clot activator eluted from plastic red-top tubes inhibited ficolin-2 ligand binding, while other related proteins, like mannose-binding lectin (MBL) and ficolin-1, were not affected. These tube types did not affect the concentrations of other related opsonins (C1q, MBL, or ficolin-3 [H-ficolin]). Interestingly, we also found that ficolin-1 levels were increased 2-to 3-fold in plastic serum separator tubes compared to the increases in other tube types. These findings have implications for future ficolin-1 and ficolin-2 studies, as proper sample collection and handling are essential.

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Section: Discussionsupporting

confidence: 92%

Blood Collection Tubes Influence Serum Ficolin-1 and Ficolin-2 Levels

Brady¹,

Spencer²,

Falsey³

et al. 2013

Clin. Vaccine Immunol.

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“…We used EDTA plasma whereas Watanabe et al used serum. It has recently been shown by us and others that Ficolin-2 binds to the coagulation activator silica applied in some serum tubes, and is thus depleted from the samples (Brady et al, 2014;Hein et al, 2013). This could be one possible explanation of the deviation between the two studies.…”

Section: Discussionmentioning

confidence: 91%

Ficolins and the lectin pathway of complement in patients with systemic lupus erythematosus

Hein

Nielsen

et al. 2015

Molecular Immunology

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“…We and others have reported the specific inhibition of ficolin-2 by the coating in plastic serum collection tubes (Brady et al, 2014c; Hein et al, 2013), and others have observed the depletion of ficolin-2 by heparin-treated cardio bypass circuits (Hein et al, 2015). Ficolin-2 was observed to bind to derivatized Sepharose (Kilpatrick & Chalmers, 2012), and commercial complement component-depleted sera are deplete of ficolin-2 as well (Brady et al, 2014b).…”

Section: Discussionmentioning

confidence: 93%

Ficolin-2 inhibitors are present in sera after prolonged storage at −80 °C

Geno

Kennedy

Sawyer

et al. 2016

PeerJ

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Ficolins can activate the lectin pathway of the complement system that provides innate immune protection against pathogens, marks host cellular debris for clearance, and promotes inflammation. Baseline inflammation increases with aging in a phenomenon known as “inflammaging.” Although IL-6 and C-reactive protein are known to increase with age, contributions of many complement factors, including ficolins, to inflammaging have been little studied. Ficolin-2 is abundant in human serum and can recognize many target structures; therefore, ficolin-2 has potential to contribute to inflammaging. We hypothesized that inflammaging would alter ficolin-2 levels among older adults and examined 360 archived sera collected from older individuals. We found that these sera had apparently reduced ficolin-2 levels and that 84.2% of archived sera exhibited ficolin-2 inhibitors, which suppressed apparent amounts of ficolin-2 detected by enzyme-linked immunosorbent assay. Fresh serum samples were obtained from donors whose archived sera showed inhibitors, but the fresh sera did not have ficolin-2 inhibitors. Ficolin-2 inhibitors were present in other long-stored sera from younger persons. Furthermore, noninhibiting samples and fresh sera from older adults had apparently normal amounts of ficolin-2. Thus, ficolin-2 inhibitors may arise as an artifact of long-term storage of serum at −80 °C.

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Ficolin-2 reveals different analytical and biological properties dependent on different sample handling procedures

Cited by 25 publications

References 32 publications

Blood Collection Tubes Influence Serum Ficolin-1 and Ficolin-2 Levels

Blood Collection Tubes Influence Serum Ficolin-1 and Ficolin-2 Levels

Ficolins and the lectin pathway of complement in patients with systemic lupus erythematosus

Ficolin-2 inhibitors are present in sera after prolonged storage at −80 °C

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