Antibodies specific for phosphorylation sites and screening methods using the same antibodies

• US 7,417,124 B2
• Filed: 04/13/2004
• Issued: 08/26/2008
• Est. Priority Date: 10/09/2003
• Status: Active Grant

• Current Assignee: University of Kansas Medical Center, Zeria Pharmaceutical Company Limited
• Original Assignee: University of Kansas Medical Center, Zeria Pharmaceutical Company Limited
• Inventors: Fujisawa, Junichi, Mori, Shigeo, Matsuzaki, Koichi, Seki, Toshihito, Yoshida, Katsunori, Nishizawa, Mikio, Furukawa, Fukiko, Matsushita, Masanori, Sugano, Yasushi, Yamagata, Hideo, Tahashi, Yoshiya
• Primary Examiner(s): Canella; Karen A.
• Assistant Examiner(s): Reddig; Peter J
• Application Number:
  US10/822,860
• Publication Number:
  US 20050079559A1
• Time in Patent Office: 1,596 Days
• Field of Search: 530/387.1 424/130.1
• US Class Current: 530/387.1
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 involving specific cell types
• CPC Class Codes: G01N 33/5067 Liver cells
• CPC Class Codes: G01N 33/5088 of vertebrates
• CPC Class Codes: G01N 33/57419 of colon

Abstract

Polyclonal antibody specific for a phosphorylated linker region in Smad2 and/or Smad3.

Claims

1. A polyclonal antibody specific for a phosphorylated linker region in Smad2, obtained from antiserum raised by immunizing a mammal with a phosphorylated product of a peptide including an amino acid sequence in the linker region of Smad2, wherein the phosphorylated product of a peptide including the amino acid sequence in the linker region of Smad2 for the immunization is:

Pro Ala Glu Leu p-Ser Pro Thr Thr Leu p-Ser Pro Val Asn His Ser (SEQ ID NO;

     1) wherein p-Ser represents phosphorylated serine.

3. The polyclonal antibody according to claims 1 or 2, wherein the mammal is a rabbit.

4. The polyclonal antibody according to claims 1 or 2, wherein the raised antiserum is affinity purified with a phosphorylated peptide(s).

2. A polyclonal antibody specific for a phosphorylated linker region in Smad3, obtained from antiserum raised by immunizing a mammal with a phosphorylated product of a peptide including an amino acid sequence in the linker region of Smad3, wherein the phosphorylated product of a peptide including the amino acid sequence of the linker region of Smad3 for the immunization is:

Ala Gly Ser Pro Asn Leu p-Ser Pro Asn Pro Met p-Ser Pro Ala (SEQ ID NO

     2) wherein p-Ser represents phosphorylated serine.