An enzyme-linked immunosorbent assay (ELISA) for quantification of human collectin 11 (CL-11, CL-K1)

L Selman, M L Henriksen, J Brandt, Y Palarasah, A Waters, P L Beales, U Holmskov, T J D Jørgensen, C Nielsen, K Skjodt, Søren Hansen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Collectin 11 (CL-11), also referred to as collectin kidney 1 (CL-K1), is a pattern recognition molecule that belongs to the collectin group of proteins involved in innate immunity. It interacts with glycoconjugates on pathogen surfaces and has been found in complex with mannose-binding lectin-associated serine protease 1 (MASP-1) and/or MASP-3 in circulation. Mutation in the CL-11 gene was recently associated with the developmental syndrome 3MC. In the present study, we established and thoroughly validated a sandwich enzyme-linked immunosorbent assay (ELISA) based on two different monoclonal antibodies. The assay is highly sensitive, specific and shows excellent quantitative characteristics such as reproducibility, dilution linearity and recovery (97.7-104%). The working range is 0.15-34 ng/ml. The CL-11 concentration in two CL-11-deficient individuals affected by the 3MC syndrome was determined to be below 2.1 ng/ml. We measured the mean serum CL-11 concentration to 284 ng/ml in 100 Danish blood donors, with a 95% confidence interval of 269-299 ng/ml. There was no significant difference in the CL-11 concentration measured in matched serum and plasma samples. Storage of samples and repeated freezing and thawing to a certain extent did not influence the ELISA. This ELISA offers a convenient and reliable method for studying CL-11 levels in relation to a variety of human diseases and syndromes.
Original languageEnglish
JournalJournal of Immunological Methods
Volume375
Issue number1-2
Pages (from-to)182-8
Number of pages7
ISSN0022-1759
DOIs
Publication statusPublished - 2012

Keywords

  • Animals
  • Antibodies, Monoclonal
  • CHO Cells
  • Cells, Cultured
  • Collectins
  • Cricetinae
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Reproducibility of Results
  • Sensitivity and Specificity

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