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Methods for detection of folate receptor 1 in a patient sample

A technology for detecting folic acid receptors and samples, applied in the direction of material inspection products, chemical instruments and methods, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve limited false positive results, do not provide Issues such as shedding FOLR1 sensitivity

Pending Publication Date: 2020-05-05
IMMUNOGEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, many current assays such as ELISA-based assays do not provide sufficient sensitivity to distinguish changes in shed FOLR1 at physiological levels and are limited by false-positive results

Method used

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  • Methods for detection of folate receptor 1 in a patient sample
  • Methods for detection of folate receptor 1 in a patient sample
  • Methods for detection of folate receptor 1 in a patient sample

Examples

Experimental program
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Effect test

Embodiment 1

[0194] sFRα Immunocapture-LC / MS Assay

[0195] An ELISA based assay was developed and described in WO 2014 / 036495 / . This assay was performed using ELISA plates coated with FOLR1-Fc (a fusion peptide of huFOLR1 and murine IgG2A hinge, CH2, CH3), muFR1-9 as capture antibody, and biotinylated muFR1-13. The assay was optimized by a systematic approach where the criteria were reproducible signal with high signal-to-noise ratio, minimal matrix effects in human plasma samples, high reproducibility and precision, and lowest detection limit. The optimal ELISA conditions chosen resulted in the detection of sFRα as low as 25 ng / mL with negligible / low noise. Despite optimized conditions, this ELISA assay did not provide sufficient sensitivity to distinguish sFRα levels in normal subjects from the distribution of sFRα levels in ovarian cancer patients to allow association analysis of sFRα levels and disease state.

[0196] A more recent ELISA-based assay using two different human FRα-spe...

Embodiment 2

[0202] Data Analysis of Human Plasma Samples Using the sFRα Immunocapture-LC / MS Assay

[0203] Experiments were designed using the above parameters to measure the levels of sFRα in samples containing low levels of sFRα at 0.3 ng / mL in the surrogate matrix. Such as figure 2 As shown in , the results of these experiments demonstrate that very low levels of sFRα can be measured by the immunocapture-LC / MS assay. Experiments were also designed to measure sFRα levels in normal human plasma samples. Such as image 3 As shown in , endogenous sFRα levels seen at approximately 0.5 ng / mL could be measured by immunocapture-LC / MS assay. Figure 4 Extracted ion chromatograms of predose samples from patients with elevated sFRα are shown. The results of this experiment demonstrated that patient plasma samples with elevated levels of sFRα can be clearly distinguished from normal human plasma samples with endogenous levels.

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Abstract

The invention generally relates to methods and kits for the detection of human folate receptor 1 in a sample. Peptides of human folate receptor 1 are further provided.

Description

technical field [0001] The field of the invention relates generally to methods and kits for detecting human folate receptor 1 (FOLR1 ) in a sample. [0002] Background of the invention [0003] Cancer is a leading cause of death in the developed world, with more than 1 million people diagnosed with cancer and more than 500,000 deaths each year in the United States alone. Overall, it is estimated that more than 1 / 3 of people will develop some form of cancer during their lifetime. There are more than 200 different types of cancer, four of which—breast, lung, colorectal, and prostate—account for more than half of all new cases (Jemal et al., 2003, Cancer J. Clin. 53:5-26). [0004] Folate receptor 1 (FOLR1), also known as folate receptor-alpha or folate-binding protein, is an N-glycosylated protein expressed on the plasma membrane of cells. FOLR1 has high affinity for folate as well as for several reduced folate derivatives. FOLR1 mediates the delivery of physiological folic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/566G01N33/68
CPCG01N33/543G01N33/6848G01N33/82C07K16/28C07K2317/565C07K7/08G01N33/54326G01N33/68C07K16/18C07K7/06G01N30/7233G01N33/577G01N2030/8831G01N33/57449C07K2317/92G01N2560/00G01N33/57484
Inventor 雷蒙德·徐凯瑞·库尔姆-默德克
Owner IMMUNOGEN INC