US2007020281A1PendingUtilityA1

Monoclonal antibodies specific for anthrax spores and peptides derived from the antibodies thereof

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Individually held — no corporate assignee on recordPriority: Apr 29, 1998Filed: Jul 1, 2005Published: Jan 25, 2007
Est. expiryApr 29, 2018(expired)· nominal 20-yr term from priority
Inventors:John F. Kearney
G01N 33/56911C07K 2317/565C07K 16/1278G01N 2333/32
40
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Claims

Abstract

The present invention relates to peptides that bind to the bacterial spores, such as B. anthracis, B. subtilis and B. cereus spores. The present invention also relates to method of identifying such peptides using phage display ligand screening system. The present invention further relates to the use of such peptides for the detection of bacterial spores, such as Bacillus anthracis spores.

Claims

exact text as granted — not AI-modified
1 . An isolated peptide which binds to a  B. anthracis  spore, said peptide comprising an amino acid sequence selected from the formula of ATYPX 3 PX 4 R (SEQ IS NO: 129) or ATYPX 3 PX 4 RGGGC (SEQ ID NO:132), wherein said X 3  is Ile, Val, Leu or Pro; X 4  is Ile, Phe, His, or Thr.  
     
     
         2 . The isolated peptide of  claim 1 , wherein said X 3  is Leu or Val, X 4  is Ile, or Thr.  
     
     
         3 . The isolated peptide of  claim 2 , wherein said X 3  is Leu and X 4  is Ile.  
     
     
         4 . An isolated peptide which binds to a  B. anthracis  spore, said peptide comprising an amino acid sequence of SEQ ID NO: 127, or SEQ ID NO: 128.  
     
     
         5 . An isolated peptide which binds to a  B. anthracis  spore, said peptide comprising an amino acid sequence selected from the formula of TYPX 1 PX 2 R (SEQ ID NO: 119), wherein said X 1  is Ile, Val, Leu or Pro; X 2  is Ile, Phe, His, or Thr.  
     
     
         6 . The isolated peptide of  claim 5 , wherein said X 1  is Val or Leu X 2  is Ile or Phe.  
     
     
         7 . An isolated peptide which binds to a  B. anthracis  spore, said peptide comprising an amino acid sequence of SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO:43, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 121, SEQ ID NO: 122, or SEQ ID NO: 123.  
     
     
         8 . The isolated peptide of  claim 7 , wherein said peptide has an amino acid sequence which is selected from the group consisting of: SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 121, SEQ ID NO: 122, and SEQ ID NO: 123.  
     
     
         9 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 40.  
     
     
         10 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 41.  
     
     
         11 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 42.  
     
     
         12 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 43.  
     
     
         13 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 121.  
     
     
         14 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 122.  
     
     
         15 . The isolated peptide of  claim 7 , wherein said peptide has the amino acid sequence of SEQ ID NO: 123.  
     
     
         16 . A method for detecting a  B. anthracis  spore in a sample, said method comprising: 
 (a) contacting said sample to a peptide which has an amino acid sequence of SEQ ID NO. 127 or SEQ ID NO. 128;    (b) determining the presence of said  B. anthracis  spores in said sample.    
     
     
         17 . The method of  claim 16 , wherein said determining the presence of  B. anthracis  spores in said sample is performed using fluorescence-activated cell sorting (FACS).  
     
     
         18 . The method of  claim 16 , wherein said determining the presence of  B. anthracis  spores in said sample is performed using fluorescence microscopy.  
     
     
         19 . A method of detecting a  B. anthracis  spore in a sample, said method comprising: 
 (a) contacting said sample to a peptide which comprises an amino acid sequence of SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 121, SEQ ID NO: 122, or SEQ ID NO: 123,    (b) determining the presence of said  B. anthracis  spores in said sample.    
     
     
         20 . The method of  claim 19 , wherein said determining the presence of  B. anthracis  spores in said sample is performed using fluorescence-activated cell sorting (FACS).  
     
     
         21 . The method of  claim 19 , wherein said determining the presence of  B. anthracis  spores in said sample is performed using fluorescence microscopy.  
     
     
         22 . A method of identifying a peptide which binds to a  B. anthracis  spore comprising the steps of: 
 a. mixing phage from a Phage Display library with said  B. anthracis  spores;    b. incubating the product of step (a) for sufficient time to allow the formation of phage-spore complexes;    c. obtaining said phage-spore complexes;    d. washing said phage-spore complexes repeatedly;    e. eluting the phage from said phage-spore complexes with elution buffer;    f. amplifying the eluted phage;    g. purifying individual clones;    h. amplifying purified clones, and then extracting genomic DNA from each preparation.    
     
     
         23 . The method of  claim 22 , following the step (e), further comprising the step of neutralizing said eluate.  
     
     
         24 . The method of  claim 22 , further comprising the step (i), determining the DNA sequence encoding peptides.  
     
     
         25 . The method of  claim 24 , following the step (i), further comprising the step of testing binding activity of said peptides to said  anthracis  spores.  
     
     
         26 . The method of  claim 22 , following the step (f), further comprising the step of repeating said steps (a)-(f) 2 to 5 rounds.  
     
     
         27 . A Kit for detecting a  B. anthracis  spore comprising a peptide of  claim 4 .  
     
     
         28 . The Kit of  claim 27  further comprising a detective tag.  
     
     
         29 . The Kit of  claim 28 , wherein said detective tag is fluorescent agent, phosphorescent agent, calorimetric agent or radioactive agent.  
     
     
         30 . A Kit for detecting a  B. anthracis  spore comprising a peptide of  claim 7 .  
     
     
         31 . The Kit of  claim 30  further comprising a detective tag.  
     
     
         32 . The Kit of  claim 31 , wherein said detective tag is fluorescent agent, phosphorescent agent, calorimetric agent or radioactive agent.

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