Assay for plasma cell associated disease

Abstract

The application provides a method of identifying or monitoring a plasma cell associated disease, comprising purifying immunoglobulin free light chains (FLCs) from a sample from a subject with anti-FLC specific antibodies or fragments thereof and subjecting the purified sample to a mass spectrometry technique to identify the presence of one or more peaks corresponding to one or more monoclonal FLCs in the sample.

Description

(19) *EP004675278A3* (11) EP 4 675 278 A3 (12) EUROPEAN PATENT APPLICATION (88) Date of publication A3: 08.04.2026 Bulletin 2026 / 15 (43) Date of publication A2: 07.01.2026 Bulletin 2026 / 02 (21) Application number: 25217662.3 (22) Date of filing: 23.05.2018 (51) International Patent Classification (IPC): G01N 33 / 68 (2006.01) G01N 33 / 57505 (2026.01) (52) Cooperative Patent Classification (CPC): G01N 33 / 6848; G01N 33 / 57505; G01N 33 / 6854; G01N 33 / 6857 (84) Designated Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR (30) Priority: 23.05.2017 GB 201708262 (62) Document number(s) of the earlier application(s) in accordance with Art. 76 EPC: 22175618.2 / 4 071 478 18728707.3 / 3 631 452 (71) Applicant: The Binding Site Group Limited Birmingham, West Midlands B15 1QT (GB) (72) Inventors: • WALLIS, Gregg Birmingham, B15 1QT (GB) • HARDING, Stephen Birmingham, B15 1QT (GB) • HUGHES, Richard Geir Birmingham, B15 1QT (GB) (74) Representative: Henderson, Helen Lee Withers & Rogers LLP 2 London Bridge London SE1 9RA (GB) (54) ASSAY FOR PLASMA CELL ASSOCIATED DISEASE (57) The application provides amethod of identifying ormonitoring a plasma cell associated disease, compris- ing purifying immunoglobulin free light chains (FLCs) from a sample from a subject with anti-FLC specific antibodies or fragments thereof and subjecting the pur- ified sample to amass spectrometry technique to identify the presence of one or more peaks corresponding to one or more monoclonal FLCs in the sample. EP 4 67 5 27 8 A 3 Processed by Luminess, 75001 PARIS (FR) 2 EP 4 675 278 A3 5 10 15 20 25 30 35 40 45 50 55 3 EP 4 675 278 A3 5 10 15 20 25 30 35 40 45 50 55 4 EP 4 675 278 A3 5 10 15 20 25 30 35 40 45 50 55 5 EP 4 675 278 A3 5 10 15 20 25 30 35 40 45 50 55 摘要 本申请提供了鉴定或监测浆细胞相关疾病的方法，其包括用抗 FLC 特异性抗体或其片段从 来自对象的样品中纯化免疫球蛋白游离轻链(FLC)，并使纯化的样品经受质谱技术以鉴定样 品中对应于一个或多个单克隆 FLC 的一个或多个峰的存在。

Claims

1. A method of identifying or monitoring a plasma cell associated disease, comprising: purifying immunoglobulin free light chains (FLCs) from a sample from a subject with: i) a mixture of anti-kappa FLC specific antibodies and anti-lambda FLC specific antibodies; ii) a mixture of anti-kappa FLC specific antibodies and anti-lambda FLC specific antibody fragments; iii) a mixture of anti-kappa FLC specific antibody fragments and anti-lambda FLC specific antibodies; or iv) a mixture of anti-kappa FLC specific antibody fragments and anti-lambda FLC specific antibody fragments; and subjecting the purified sample to a mass spectrometry technique to identify the presence of one or more peaks corresponding to one or more monoclonal FLCs in the sample2. A method according to claim 1, wherein the anti-kappa FLC specific antibodies, the anti-lambda FLC specific antibodies. the anti-kappa FLC specific antibody fragments, or the anti-lambda FLC specific antibody fragments are polyclonal.

3. A method according to claims 1 or 2, wherein the anti-kappa FLC specific antibody fragments, or the anti-lambda FLC specific antibody fragments are F(ab')2 fragments.

4. A method according to any of claims 1 to 3, wherein the anti-FLC kappa specific antibodies, or the anti-lambda FLC specific antibodies, comprise one or more non-disulphide cross-links between at least one heavy chain and one light chain of the antibody5. A method according to any of claims 1 to 4, wherein the anti-kappa FLC specific antibody fragments, or the anti-lambda FLC specific antibody fragments comprise one or more non-disulphide cross-links between at least one heavy chain fragment and one light chain fragment of the fragment of the antibody.

6. A method according to any of claims 1 to 5, wherein the mass spectrometry technique is an Orbitrap mass spectrometer, ion trap mass spectrometer, time-of-flight mass spectrometer, triple quadrupole mass spectrometer, or quadrupole mass spectrometer may be used.

7. A method of claim 6, wherein the mass spectrometry techniques is matrix assisted laser desorption ionisation-time-of-flight mass spectrometry (MALDI-TOF)8. A method according to any preceding claim, additionally comprising: purifying total kappa and total lambda light chains with: i) a mixture of anti-kappa specific antibodies and anti-lambda specific antibodies; ii) a mixture of anti-kappa specific antibodies and anti-lambda specific antibody fragments; iii) a mixture of anti-kappa specific antibody fragments and anti-lambda specific antibodies; or iv) a mixture of anti-kappa specific antibody fragments and anti-lambda specific antibody fragments; and subjecting the purified sample to mass spectrometry, to identify one or more peaks corresponding to one or more monoclonal immunoglobulins.

9. A method according to any of claims 1 to 8, wherein the plasma cell associated disease is selected from intact immunoglobulin, multiple myeloma, light chain multiple myeloma, non-secretory multiple myeloma, AL amyloidosis, light chain deposition disease (LCDD), smouldering multiple myeloma, monoclonal gammopathy of undetermined significance (MGUS), macroglobulinemia, POEMS (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy and skin changes) syndrome and LCDD.

10. A method according to claims any of 1 to 9, wherein the sample is tear fluid, plasma, serum, saliva, urine, blood or cerebrospinal fluid.

Images