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Circulating Levels of sFlt1 Splice Variants as Predictive Markers for the Development of Preeclampsia.

Souders CA, Maynard SE, Yan J, Wang Y, Boatright NK, Sedan J, Balyozian D, Cheslock PS, Molrine DC, Simas TA - Int J Mol Sci (2015)

Bottom Line: Here we describe the development and use of diagnostic monoclonal antibodies specific to the two main splice variants of sFlt1, sFlt1-1 and sFlt1-14.These antibodies were selected for their sensitivity and specificity to their respective sFlt1 isoform in a capture ELISA format.It may be possible to improve current diagnostic platforms if more specific antibodies are utilized.

View Article: PubMed Central - PubMed

Affiliation: MassBiologics of the University of Massachusetts Medical School, Boston, MA 02126, USA. Colby.Souders@UMassMed.edu.

ABSTRACT
Angiogenic biomarkers, including soluble fms-like tyrosine kinase 1 (sFlt1), are thought to be predictors of preeclampsia onset; however, improvement is needed before a widespread diagnostic test can be utilized. Here we describe the development and use of diagnostic monoclonal antibodies specific to the two main splice variants of sFlt1, sFlt1-1 and sFlt1-14. These antibodies were selected for their sensitivity and specificity to their respective sFlt1 isoform in a capture ELISA format. Data from this pilot study suggest that sFlt1-1 may be more predictive of preeclampsia than total sFlt1. It may be possible to improve current diagnostic platforms if more specific antibodies are utilized.

No MeSH data available.


Related in: MedlinePlus

Mouse mAbs Ex14-1 (A) and 1CKLH18 (B) specifically detected endogenous sFlt1 isoforms from amniotic fluid on a Western blot. Each mAb recognized a single protein of the expected molecular weight (~115 kDa) when compared to recombinant standards. Included as a positive control (C) is a commercially available mAb (Sigma Cat.#V4262) that recognized total sFlt1.
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ijms-16-12436-f004: Mouse mAbs Ex14-1 (A) and 1CKLH18 (B) specifically detected endogenous sFlt1 isoforms from amniotic fluid on a Western blot. Each mAb recognized a single protein of the expected molecular weight (~115 kDa) when compared to recombinant standards. Included as a positive control (C) is a commercially available mAb (Sigma Cat.#V4262) that recognized total sFlt1.

Mentions: Specificity of mAbs to sFlt1-14 and sFlt1-1 was further investigated by Western blot analysis on human amniotic fluid (Figure 4). Both Ex14-1 (Figure 4A) and 1CKLH18 (Figure 4B) recognized a single protein in amniotic fluid at the expected molecular weight (~115 kDa) that corresponds to recombinant human sFlt1-14 and sFlt1-1, respectively. In addition, both mouse mAbs specifically recognized their recombinant sFlt1 isoform standards by Western blot. These data confirm the ELISA results suggesting the mouse antibodies were specific for their sFlt1 isoforms and recognized their endogenous sFlt1 isoform in biological fluid.


Circulating Levels of sFlt1 Splice Variants as Predictive Markers for the Development of Preeclampsia.

Souders CA, Maynard SE, Yan J, Wang Y, Boatright NK, Sedan J, Balyozian D, Cheslock PS, Molrine DC, Simas TA - Int J Mol Sci (2015)

Mouse mAbs Ex14-1 (A) and 1CKLH18 (B) specifically detected endogenous sFlt1 isoforms from amniotic fluid on a Western blot. Each mAb recognized a single protein of the expected molecular weight (~115 kDa) when compared to recombinant standards. Included as a positive control (C) is a commercially available mAb (Sigma Cat.#V4262) that recognized total sFlt1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490453&req=5

ijms-16-12436-f004: Mouse mAbs Ex14-1 (A) and 1CKLH18 (B) specifically detected endogenous sFlt1 isoforms from amniotic fluid on a Western blot. Each mAb recognized a single protein of the expected molecular weight (~115 kDa) when compared to recombinant standards. Included as a positive control (C) is a commercially available mAb (Sigma Cat.#V4262) that recognized total sFlt1.
Mentions: Specificity of mAbs to sFlt1-14 and sFlt1-1 was further investigated by Western blot analysis on human amniotic fluid (Figure 4). Both Ex14-1 (Figure 4A) and 1CKLH18 (Figure 4B) recognized a single protein in amniotic fluid at the expected molecular weight (~115 kDa) that corresponds to recombinant human sFlt1-14 and sFlt1-1, respectively. In addition, both mouse mAbs specifically recognized their recombinant sFlt1 isoform standards by Western blot. These data confirm the ELISA results suggesting the mouse antibodies were specific for their sFlt1 isoforms and recognized their endogenous sFlt1 isoform in biological fluid.

Bottom Line: Here we describe the development and use of diagnostic monoclonal antibodies specific to the two main splice variants of sFlt1, sFlt1-1 and sFlt1-14.These antibodies were selected for their sensitivity and specificity to their respective sFlt1 isoform in a capture ELISA format.It may be possible to improve current diagnostic platforms if more specific antibodies are utilized.

View Article: PubMed Central - PubMed

Affiliation: MassBiologics of the University of Massachusetts Medical School, Boston, MA 02126, USA. Colby.Souders@UMassMed.edu.

ABSTRACT
Angiogenic biomarkers, including soluble fms-like tyrosine kinase 1 (sFlt1), are thought to be predictors of preeclampsia onset; however, improvement is needed before a widespread diagnostic test can be utilized. Here we describe the development and use of diagnostic monoclonal antibodies specific to the two main splice variants of sFlt1, sFlt1-1 and sFlt1-14. These antibodies were selected for their sensitivity and specificity to their respective sFlt1 isoform in a capture ELISA format. Data from this pilot study suggest that sFlt1-1 may be more predictive of preeclampsia than total sFlt1. It may be possible to improve current diagnostic platforms if more specific antibodies are utilized.

No MeSH data available.


Related in: MedlinePlus