Compositions and methods for apo-b48 and apo-b100 assay

• US 20050152900A1
• Filed: 06/19/2003
• Published: 07/14/2005
• Est. Priority Date: 06/19/2002
• Status: Abandoned Application

• Current Assignee: Genfit SA
• Original Assignee: Genfit SA
• Inventors: Najib, Jamila, Majd, Zouher
• Application Number:
  US10/513,916
• Publication Number:
  US 20050152900A1
• Time in Patent Office: Days
• US Class Current: 424/145.100
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/775 Apolipopeptides
• CPC Class Codes: C07K 16/18 against material from animals or humans

Abstract

The invention concerns compositions and methods for assay or detection of apolipoprotein-B48 in samples. In particular, it concerns a method for differential measurement of apolipoprotein-B48 (“Apo-B48”) and apolipoprotein-B100 (“Apo-B100”) in biological samples. The invention also concerns synthetic products of Apo-B100, the corresponding antibodies, kits containing same, and their uses for detecting, differentially quantifying and/or recording an amount of Apo-B48 and/or Apo-B100 in a sample, or for quantifying and/or recording atherogenic lipoparticles in a sample. The products, materials and kits hereinabove can also be used for differentially modulating the levels of Apo-B48 and/or Apo-B100 or their activity, in vitro or in vivo, and for regulating lipid metabolism in a subject.

Claims

1-33. -33. (canceled)

34. A substantially pure synthetic peptide, comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2 or an immunogenic fragment or a derivative of said sequences.

35. A substantially pure synthetic peptide according to claim 34, comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2.

36. The peptide according to claim 34, wherein the peptide is soluble or complexed with a carrier molecule such as KLH, serum albumin or a bead.

37. The peptide according to claim 34, wherein it allows the production of ApoB100-immunoprecipitant antibodies.

43. A method for producing a specific anti-Apo-B100 antibody, comprising administering a peptide according to claim 34 or a mixture thereof to a non-human animal and recovering the antibodies or antibody-producing cells.

44. A method for detecting or quantifying Apo-B100 in a biological sample, comprising contacting the sample with an antibody specific to a peptide according to claim 34 or with an antibody produced by a method according to claim 43, and detecting the presence of antigen-antibody immune complexes.

45. A method for detecting or differentially quantifying Apo-B100 and Apo-B48 in a sample, comprising the following steps:

(a) determining the amount of total Apo-B in a sample with a total anti-Apo-B antibody,

(b) eliminating Apo-B100 or lipoparticles containing Apo-B100 from said sample, with the help of a specific anti-Apo-B100 antibody specific of a peptide according to claim 34 or produced by a method according to claim 43, and

(c) determining the amount of Apo-B48 in the sample obtained after step (b) or in lipoparticles not containing Apo-B100 in said sample, with a total anti-Apo-B antibody.

46. The method according to claim 45, wherein, during steps (a) and/or (c), a polyclonal antibody directed against total anti-Apo-B is used.

47. The method according to claim 45, wherein the specific anti-Apo-B100 antibody used in step (b) is a polyclonal antibody or a mixture of monoclonal antibodies or a modified antibody allowing precipitation of immune complexes.

48. The method according to claim 45, wherein the specific anti-Apo-B100 antibody used in step (b) is an immunoprecipitant polyclonal antibody.

49. The method according to claim 45, wherein the presence of an antigen-antibody immune complex is determined by ELISA, RIA, EIA, turbidimetry or any other immunological assay.

50. The method according to claim 45, wherein the presence of an antigen-antibody immune complex is determined by a nephelometric analytical method.

51. A method for detecting or differentially quantifying Apo-B100 and Apo-B48 in a biological sample comprising contacting said sample with an antibody specific of a peptide according to claim 34 or with an antibody produced by a method according to claim 43, and indirectly detecting the formation of Apo-B100-antibody immune complexes by a nephelometric analytical method.

52. The method according to claim 45, wherein the biological sample is a sample of blood or plasma or serum or interstitial fluid or cerebrospinal fluid or a cell culture supernatant.

53. A method for detecting the presence, a predisposition or a risk of developing a disorder of lipid metabolism in a subject, comprising differentially quantifying in vitro the amounts of Apo-B48 and Apo-B100 in a sample from a subject, with an antibody specific of a peptide according to claim 34, or with an antibody produced by a method according to claim 43.

54. The method according to claim 53, for detecting the presence, a predisposition or a risk of developing atherosclerosis or a cardiovascular disease such as a coronary disease for example.

55. A method for monitoring a treatment for correcting disorders of lipid metabolism in a subject, comprising quantifying Apo-B100 and/or Apo-B48 in vitro, in a sample from said subject, with the help of an antibody specific of a peptide according to claim 34 or an antibody produced by a method according to claim 43.

56. The method according to claim 55, for monitoring a treatment of atherosclerosis or of a cardiovascular disease such as a coronary disease for example.

57. A method for detecting or monitoring the cellular uptake of LDL and triglyceride-rich lipoproteins in a subject, comprising detecting in vitro the amounts of Apo-B48 and/or Apo-B100 present in lipoparticles, with the help of an antibody specific of a peptide according to claim 34 or an antibody produced by a method according to claim 43.

58. A method for evaluating the physiological state of a subject, comprising detecting the levels of Apo-B48 and Apo-B100 in vitro in a sample from said subject, with the help of an antibody specific of a peptide according to claim 34 or an antibody produced by a method according to claim 43.

59. A method for screening compounds or diets capable of modulating the serum or plasma concentration of Apo-B48 and/or Apo-B100, by implementing an antibody specific of a peptide according to claim 34 or produced by a method according to claim 43.

60. The antibody specific of a peptide according to claim 34 or produced by a method according to claim 43, wherein the antibody is coupled with heterologous fragments such as toxins, labels, medicaments or any other therapeutic agent.

61. A method for selectively regulating Apo-B100 activity in vivo, by administering into a subject in need of such treatment an antibody specific of a peptide according to claim 34 or produced by a method according to claim 43.

62. A pharmaceutical composition comprising an antibody specific of a peptide, comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2 or an immunogenic fragment or a derivative of said sequences, or an antibody produced by a method according to claim 43, and a pharmaceutically acceptable vehicle or excipient.

65. A kit comprising a substantially pure synthetic peptide comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2 or an immunogenic fragment or a derivative of said sequences, or an antibody specific of said peptide or an antibody produced by a method according to claim 43.

38. An antibody specific of a synthetic peptide, comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2 or an immunogenic fragment or a derivative of said sequences, or fragment or derivative of said antibody having substantially the same antigenic specificity.

39. The antibody according to claim 38, wherein it is produced by immunizing a non-human animal with the peptide, and recovering the antibodies or the antibody-producing cells.

40. The antibody according to claim 38, wherein it is a polyclonal antibody.

41. The antibody according to claim 38, wherein it corresponds to a mixture of monoclonal antibodies.

42. The antibody according to claim 38, wherein it is capable of precipitating Apo-B100.

63. A method for screening compounds modulating Apo-B activity by implementing a substantially pure synthetic peptide, comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2 or an immunogenic fragment or a derivative of said sequences.

64. A method for specifically modulating Apo-B100 activity, by implementing a substantially pure synthetic peptide comprising the amino acid sequence SEQ ID NO:

1 or SEQ ID NO;

2 or an immunogenic fragment or a derivative of said sequences.