2015
DOI: 10.1371/journal.pone.0116237
|View full text |Cite
|
Sign up to set email alerts
|

Anti-Lubricin Monoclonal Antibodies Created Using Lubricin-Knockout Mice Immunodetect Lubricin in Several Species and in Patients with Healthy and Diseased Joints

Abstract: Lubricin, encoded by the gene PRG4, is the principal lubricant in articulating joints. We immunized mice genetically deficient for lubricin (Prg4-/-) with purified human lubricin, and generated several mAbs. We determined each mAb’s binding epitope, sensitivity, and specificity using biologic samples and recombinant lubricin sub-domains, and we also developed a competition ELISA assay to measure lubricin in synovial fluid and blood. We found the mAbs all recognized epitopes containing O-linked oligosaccharides… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
2
2
1

Citation Types

1
43
0

Year Published

2015
2015
2023
2023

Publication Types

Select...
9

Relationship

5
4

Authors

Journals

citations
Cited by 42 publications
(44 citation statements)
references
References 33 publications
1
43
0
Order By: Relevance
“…2 A standard concentration curve was constructed by diluting a concentrated rhPRG4 solution as we did not have purified porcine PRG4 available.…”
Section: Methodsmentioning
confidence: 99%
“…2 A standard concentration curve was constructed by diluting a concentrated rhPRG4 solution as we did not have purified porcine PRG4 available.…”
Section: Methodsmentioning
confidence: 99%
“…Both osteochondral and synovial membrane sections were incubated with 10% hyaluronidase solution for 30 min at 37°C prior to rinsing and quenching endogenous peroxidase activity for immunohistochemistry. Histological staining using lubricin-specific monoclonal antibody 6a8 (courtesy of G Jay) at a 1:1000 dilution was performed using a similar protocol to that described for anti-lubricin mAb-7h12, beginning with the quenching of endogenous peroxidase activity 28 . Safranin O/Fast Green staining was performed to assess cartilage sGAG content.…”
Section: Methodsmentioning
confidence: 99%
“…Investigators in future studies should include protein expression of chondrogenic and non-chondrogenic markers at the protein levels. This can be done by utilizing enzyme-linked immunosorbent assays (ELISAs) to measure ACAN , and VEGF [121], western blots to measure SOX9 [54], Col XI [122], lubricin [123] and osterix [124], a chloramine-T hydroxyproline assay to measure total collagen content, immunohistochemistry to measure collagen types I and X [21], and the QuantiChrom™ ALP Assay Kit to measure ALP. Of course any single one of these ancillary studies is an undertaking in itself and the authors recommend a series of careful studies to further clarify the overall chondrogenic picture.…”
Section: Discussion and Future Studiesmentioning
confidence: 99%