Binding polypeptides for B lymphocyte stimulator protein (BLyS)

• US 7,118,872 B2
• Filed: 08/17/2001
• Issued: 10/10/2006
• Est. Priority Date: 08/18/2000
• Status: Expired due to Term

• Current Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Original Assignee: Dyax Corporation (Shire plc)
• Inventors: Ladner, Robert Charles, Beltzer, James P., Fleming, Tony J.
• Primary Examiner(s): CHISM, BILLY D
• Application Number:
  US09/932,322
• Publication Number:
  US 20030194743A1
• Time in Patent Office: 1,880 Days
• Field of Search: 435/6, 435/7.1, 435/235.1, 530/327, 530/328, 536/23.1, 514/2, 514/17, 514/16, 514/13
• US Class Current: 435/7.1
• CPC Class Codes: C07K 1/047 Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 5/1021 with the first amino acid being acidic
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors

Abstract

Binding polypeptides comprising specific amino acid sequences are disclosed that bind B Lymphocyte Stimulator (BLyS) protein or BLyS-like polypeptides. The binding polypeptides can be used in methods of the invention for detecting or isolating BLyS protein or BLyS-like polypeptides in solutions or mixtures, such as blood, tissue samples, or conditioned media.

Claims

1. A B lymphocyte stimulator protein (BLyS) binding polypeptide comprising an amino acid sequence of the following formula:

Cys-X₅-Phe-X₇-Trp-Glu-Cys (residue 4–

10 of SEQ ID NO;

1), 

 

(H)

wherein

X₅ is Phe, Trp, or Tyr; and

X₇ is Pro or Tyr;

or

Cys-X₂-X₃-X₄-X₅-X₆-X₇-Cys (SEQ ID NO;

9), 

 

(I)

wherein

X₂ is Asp, Ile, Leu, or Tyr;

X₃ is Arg, Asp, Glu, His, Ile, Leu, Lys, Phe, Pro, Tyr, or Val;

X₄ is His, Leu, Lys, or Phe;

X₅ is Leu, Pro, or Thr;

X₆ is Arg, Asn, Gly, His, Ile, Lys, Met, or Trp; and

X₇ is Ala, Asn, Gln, Glu, Gly, His, Ile, Leu, Met, Phe, Ser, Trp, Tyr, or Val;

or

Cys-X₂-X₃-X₄-X₅-X₆-X₇-X₈-Cys (SEQ ID NO;

10), 

 

(J)

wherein

X₂ is Asn, or Asp;

X₃ is Arg, Asp, Ile, Leu, Met, Pro, or Val;

X₄ is Ala, Ile, Leu, Pro, Thr, or Val;

X₅ is Asn, His, Ile, Leu, Lys, Phe, or Thr;

X₆ is Asn, Glu, Gly, His, Leu, Lys, Met, Pro, or Thr;

X₇ is Arg, Asn, Asp, Gln, Glu, Gly, Ile, Lys, Met, Pro, Ser, or Trp; and

X₈ is Arg, Glu, Gly, Lys, Phe, Ser, Trp, or Tyr;

or

Cys-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-Cys (SEQ ID NO;

11), 

 

(K)

wherein

X₂ is Asp, Gln, His, Ile, Leu, Lys, Met, Phe, or Thr;

X₃ is His, Ile, Leu, Met, Phe, Pro, Trp, or Tyr;

X₄ is Asp, His, Leu, or Ser;

X₅ is Ala, Arg, Asp, Glu, Leu, Phe, Pro, or Thr;

X₆ is Ala, Arg, Asn, or Leu;

X₇ is Ile, Leu, Met, Pro, Ser, or Thr;

X₈ is Ala, Arg, Asn, Gly, His, Lys, Ser, or Tyr; and

X₉ is Ala, Arg, Asn, Gln, Leu, Met, Ser, Trp, Tyr, or Val;

or

Cys-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-Cys (SEQ ID NO;

12), 

 

(L)

wherein

X₂ is Arg, Asn, Gln, Glu, His, Leu, Phe, Pro, Trp, Tyr, or Val;

X₃ is Arg, Asp, Gln, Gly, Ile, Lys, Phe, Thr, Trp or Tyr;

X₄ is Ala, Arg, Asp, Glu, Gly, Leu, Ser, or Tyr;

X₅ is Asp, Gln, Glu, Leu, Met, Phe, Pro, Ser, or Tyr;

X₆ is Asp, Leu, Pro, Thr, or Val;

X₇ is Arg, Gln, His, Ile, Leu, Lys, Met, Phe, Thr, Trp or Tyr;

X₈ is Ala, Arg, Asn, Gln, Glu, His, Leu, Lys, Met, or Thr;

X₉ is Ala, Asn, Gln, Gly, Leu, Lys, Phe, Pro, Thr, Trp, or Tyr;

X₁₀ is Ala, Arg, Gln, His, Lys, Met, Phe, Pro, Thr, Trp, or Tyr; and

X₁₁ is Arg, Gln, Glu, Gly, His, Leu, Met, Phe, Pro, Ser, Thr, Tyr, or Val.

2. The polypeptide according to claim 1, wherein

(a) said polypeptide comprises an amino acid sequence of the formula:

Cys-X₅-Phe-X₇-Trp-Glu-Cys (residues 4–

10 SEQ ID NO;

1), and the following amino acid positions are independently selected as follows;

X₂ is Tyr;

X₄ is Pro;

or combinations of such selections;

or

(b) said polypeptide comprises an amino acid sequence of the following formula;

Cys-X₂-X₃-X₄-X₅-X₆-X₇-Cys (SEQ ID NO;

9), and the following amino acid positions are independently selected as follows;

X₂ is Asp or Leu;

X₃ is Glu or Leu;

X₄ is His or Leu;

X₅ is Thr or Pro;

X₆ is Lys;

or combinations of such selections;

or

(c) said polypeptide comprises an amino acid sequence of the following formula;

Cys-X₂-X₃-X₄-X₅-X₆-X₇-X₈-Cys (SEQ ID NO;

10), and the following amino acid positions are independently selected as follows;

X₂ is Asp;

X₃ is Ile;

X₄ is Val or Leu;

X₅ is Thr;

X₆ is Leu;

X₈ is Ser;

or combinations of such selections;

or

(d) said polypeptide comprises an amino acid sequence of the following formula;

Cys-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-Cys (SEQ ID NO;

11), and the following amino acid positions are independently selected as follows;

X₄ is Asp;

X₅ is Glu or Pro;

X₆ is Leu;

X₇ is Thr;

or combinations of such selections;

or

(e) said polypeptide comprises an amino acid sequence of the following formula;

Cys-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-Cys (SEQ ID NO;

12), and the following amino acid positions are independently selected as follows;

X₂ is Trp, Tyr, or Val;

X₃ is Asp;

X₄ is Asp;

X₅ is Leu;

X₆ is Leu or Thr;

X₇ is Lys or Thr;

X₈ is Arg or Leu;

X₉ is Thr or Trp;

X₁₀ is Met or Phe;

X₁₁ is Val;

or combinations of such selections.

3. A BLyS binding polypeptide comprising an amino acid sequence of the following formula:

(A) X₁-X₂-X₃-Cys-X₅-Phe-X₇-Trp-Glu-Cys-X₁₁-X₁₂-X₁₃ (SEQ ID NO;

1), 

 

(A)

wherein

X₁ is Ala, Asn, Lys, or Ser;

X₂ is Ala, Glu, Met, Ser, or Val;

X₃ is Ala, Asn, Lys, or Pro;

X₅ is Phe, Trp, or Tyr;

X₇ is Pro or Tyr;

X₁₁ is Ala, Gln, His, Phe, or Val;

X₁₂ is Asn, Gln, Gly, His, Ser, or Val; and

X₁₃ is Ala, Asn, Gly, Ile, Pro, or Ser;

or

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-Cys-X₁₂-X₁₃-X₁₄ (SEQ ID NO;

2), 

 

(B)

wherein

X₁ is Ala, Asp, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, Val, or is absent;

X₂ is Ala, Asn, Asp, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val;

X₃ is Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val;

X₅ is Asp, Ile, Leu, or Tyr;

X₆ is Arg, Asp, Glu, His, Ile, Leu, Lys, Phe, Pro, Tyr, or Val;

X₇ is His, Leu, Lys, or Phe;

X₈ is Leu, Pro, or Thr;

X₉ is Arg, Asn, Gly, His, Ile, Lys, Met, or Trp;

X₁₀ is Ala, Gln, Glu, Gly, His, Ile, Leu, Met, Phe, Ser, Trp, Tyr, or Val;

X₁₂ is Asp, Gln, Glu, Gly, Ile, Leu, Lys, Phe, Ser, Trp, Tyr, or Val;

X₁₃ is Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val; and

X₁₄ is Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Ile, Leu, Lys, Phe, Pro, Trp, Tyr, Val, or is absent;

or

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-Cys-X₁₃-X₁₄-X₁₅ (SEQ ID NO;

3), 

 

(C)

wherein

X₁ is Ala, Arg, Asn, Asp, Leu, Lys, Phe, Pro, Ser, or Thr;

X₂ is Asn, Asp, Gln, His, Ile, Lys, Pro, Thr, or Trp;

X₃ is Ala, Arg, Asn, Gln, Glu, His, Phe, Pro, or Thr;

X₅ is Asn, Asp, Pro, Ser, or Thr;

X₆ is Arg, Asp, Ile, Leu, Met, Pro, or Val;

X₇ is Ala, Ile, Leu, Pro, Thr, or Val;

X₈ is Asn, His, Ile, Leu, Lys, Phe, or Thr;

X₉ is Asn, Glu, Gly, His, Leu, Lys, Met, Pro, or Thr;

X₁₀ is Arg, Asn, Asp, Gln, Glu, Gly, Ile, Lys, Met, Pro, Ser, or Trp;

X₁₁ is Arg, Glu, Gly, Lys, Phe, Ser, Trp, or Tyr;

X₁₃ is Gln, Glu, Ile, Leu, Phe, Pro, Ser, Tyr, or Val;

X₁₄ is Asn, Gly, Ile, Phe, Pro, Thr, Trp, or Tyr; and

X₁₅ is Asn, Asp, Glu, Leu, Lys, Met, Pro, or Thr;

or

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-Cys-X₁₄-X₁₅-X₁₆ (SEQ ID NO;

4), 

 

(D)

wherein

X₁ is Asn, Asp, His, Leu, Phe, Pro, Ser, Tyr, or is absent;

X₂ is Arg, Asn, Asp, His, Phe, Ser, or Trp;

X₃ is Asn, Asp, Leu, Pro, Ser, or Val;

X₅ is Asp, Gln, His, Ile, Leu, Lys, Met, Phe, or Thr;

X₆ is His, Ile, Leu, Met, Phe, Pro, Trp, or Tyr;

X₇ is Asp, His, Leu, or Ser;

X₈ is Ala, Arg, Asp, Glu, Leu, Phe, Pro, or Thr;

X₉ is Ala, Arg, Asn, or Leu;

X₁₀ is Ile, Leu, Met, Pro, Ser, or Thr;

X₁₁ is Ala, Arg, Asn, Gly, His, Lys, Ser, or Tyr;

X₁₂ is Ala, Arg, Asn, Gln, Leu, Met, Ser, Trp, Tyr, or Val;

X₁₄ is Asp, Gly, Leu, Phe, Tyr, or Val; and

X₁₅ is Asn, His, Leu, Pro, or Tyr; and

X₁₆ is Asn, Asp, His, Phe, Ser, or Tyr;

or

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-Cys-X₁₆-X₁₇-X₁₈ (SEQ ID NO;

5), 

 

(E)

wherein

X₁ is Arg, Asp, Gly, His, Leu, Phe, Pro, Ser, Trp, Tyr, or is absent;

X₂ is Ala, Arg, Asn, Asp, Gly, Pro, Ser, or is absent;

X₃ is Arg, Asn, Gln, Glu, Gly, Lys, Met, Pro, Trp or Val;

X₅ is Arg, Asn, Gln, Glu, His, Leu, Phe, Pro, Trp, Tyr, or Val;

X₆ is Arg, Asp, Gln, Gly, Ile, Lys, Phe, Thr, Trp or Tyr;

X₇ is Ala, Arg, Asp, Glu, Gly, Leu, Ser, or Tyr;

X₈ is Asp, Gln, Glu, Leu, Met, Phe, Pro, Ser, or Tyr;

X₉ is Asp, Leu, Pro, Thr, or Val;

X₁₀ is Arg, Gln, His, Ile, Leu, Lys, Met, Phe, Thr, Trp or Tyr;

X₁₁ is Ala, Arg, Asn, Gln, Glu, His, Leu, Lys, Met, or Thr;

X₁₂ is Ala, Asn, Gln, Gly, Leu, Lys, Phe, Pro, Thr, Trp, or Tyr;

X₁₃ is Ala, Arg, Gln, His, Lys, Met, Phe, Pro, Thr, Trp, or Tyr;

X₁₄ is Arg, Gln, Glu, Gly, His, Leu, Met, Phe, Pro, Ser, Thr, Tyr, or Val;

X₁₆ is Arg, Asp, Gly, His, Lys, Met, Phe, Pro, Ser, or Trp;

X₁₇ is Arg, Asn, Asp, Gly, His, Phe, Pro, Ser, Trp or Tyr; and

X₁₈ is Ala, Arg, Asn, Asp, His, Leu, Phe, or Trp;

or

X₁-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂ (SEQ ID NO;

6), 

 

(F)

wherein

X₁ is Ala, Arg, Gly, His, Leu, Lys, Met, Phe, Trp, Tyr, or Val;

X₂ is Ala, Arg, Gln, His, Ile, Leu, Phe, Thr, Trp, or Tyr;

X₃ is Ala, Asp, Lys, Phe, Thr, Trp or Tyr;

X₄ is Arg, Asp, Gln, Lys, Met, Phe, Pro, Ser, Tyr, or Val;

X₅ is Asp, Leu, Lys, Phe, Pro, Ser, or Val;

X₆ is His, Ile, Leu, Pro, Ser, or Thr;

X₇ is Arg, Gly, His, Leu, Lys, Met, or Thr;

X₈ is Ala, Arg, Asn, Ile, Leu, Lys, Met, or Thr;

X₉ is Ala, Asn, Arg, Asp, Glu, Gly, His, Leu, Met, Ser, Trp, Tyr, or Val;

X₁₀ is Ile, Leu, Phe, Ser, Thr, Trp, Tyr, or Val;

X₁₁ is Ala, Arg, Gly, His, Ile, Leu, Lys, Pro, Ser, Thr, Trp, Tyr, or Val; and

X₁₂ is Arg, Asp, His, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val;

or

X₁-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃ (SEQ ID NO;

7), 

 

(G)

wherein

X₁ is Asp, Gln, Glu, Gly, His, Lys, Met, or Trp;

X₂ is Arg, Gln, His, Ile, Leu, or Pro;

X₃ is Asp, Gly, Ile, Lys, Thr, Tyr or Val;

X₄ is Asn, Asp, Gln, Glu, Met, Pro, Ser, or Tyr;

X₅ is Asn, Asp, His, Ile, Leu, Met, Pro, Thr or Val;

X₆ is Asp, Glu, His, Leu, Lys, Pro, or Val;

X₇ is Arg, Asn, Gln, His, Ile, Leu, Met, Pro, or Thr;

X₈ is Gln, Gly, His, Leu, Met, Ser, or Thr;

X₉ is Asn, Gln, Gly, His, Leu, Lys, Ser, or Thr;

X₁₀ is Ala, Gly, Ile, Leu, Lys, Met, or Phe;

X₁₁ is Ala, Glu, His, Ile, Leu, Met, Ser, Thr, Trp, Tyr, or Val;

X₁₂ is Arg, Gln, Glu, Gly, His, Ile, Lys, Tyr, or Val; and

X₁₃ is Arg, Asn, Glu, His, Ile, Ser, Thr, Trp, or Val.

4. The BLyS binding polypeptide according to claim 3, wherein

(a) said polypeptide includes an amino acid sequence of the following formula:

X₁-X₂-X₃-Cys-X₅-Phe-X₇-Trp-Glu-Cys-X₁₁-X₁₂-X₁₃ (SEQ ID NO;

1), and the following amino acid positions are independently selected as follows;

X₃ is Lys;

X₅ is Tyr;

X₇ is Pro;

X₁₁ is Ala, Gln, His, Phe, or Val;

X₁₂ is Asn, Gln, Gly, His, Ser, or Val;

X₁₃ is Ala, Asn, Gly, Ile, Pro, or Ser;

or combinations of such selections;

or

(b) said polypeptide includes an amino acid sequence of the following formula;

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-Cys-X₁₂-X₁₃-X₁₄ (SEQ ID NO;

2), and the following amino acid positions are independently selected as follows;

X₃ is Asp;

X₅ is Asp or Leu;

X₆ is Glu or Leu;

X₇ is His or Leu;

X₈ is Thr or Pro;

X₉ is Lys;

or combinations of such selections;

or

(c) said polypeptide includes an amino acid sequence of the following formula;

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-Cys-X₁₃-X₁₄-X₁₅ (SEQ ID NO;

3), and the following amino acid positions are independently selected as follows;

X₃ is Ala;

X₅ is Asp;

X₆ is Ile;

X₇ is Val or Leu;

X₈is Thr;

X₉ is Leu;

X₁₁ is Ser;

X₁₃ is Val;

X₁₅ is Glu or Pro;

or combinations of such selections;

or

(d) said polypeptide includes an amino acid sequence of the following formula;

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-Cys-X₁₄-X₁₅-X₁₆ (SEQ ID NO;

4), and the following amino acid positions are independently selected as follows;

X₁ is Ser;

X₂ is Arg;

X₃ is Asn or Asp;

X₇ is Asp;

X₈ is Glu or Pro;

X₉ is Leu;

X₁₀ is Thr;

X₁₄ is Leu;

X₁₅ is His, Leu, or Pro;

X₁₆ is Asp or Ser;

or combinations of such selections;

or

(e) said polypeptide includes an amino acid sequence of the following formula;

X₁-X₂-X₃-Cys-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃-X₁₄-Cys-X₁₆-X₁₇-X₁₈ (SEQ ID NO;

5), and the following amino acid positions are independently selected as follows;

X₁ is Arg;

X₂ is Asn, Asp, Gly, or Pro;

X₃ is Gly or Met;

X₅ is Trp, Tyr, or Val;

X₆ is Asp;

X₇ is Asp;

X₈ is Leu;

X₉ is Leu or Thr;

X₁₀ is Lys or Thr;

X₁₁ is Arg or Leu;

X₁₂ is Thr or Trp;

X₁₃ is Met or Phe;

X₁₄ is Val;

X₁₆ is Met;

X₁₇ is Arg, His, or Tyr;

X₁₈ is Asn or His;

or combinations of such selections;

or

(f) said polypeptide includes an amino acid sequence of the following formula;

X₁-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂ (SEQ ID NO;

6), and the following amino acid positions are independently selected as follows;

X₁ is Gly, Tyr, or Val;

X₂ is His or Tyr;

X₃ is Asp or Tyr;

X₄ is Asp or Gln;

X₅ is Leu or Ser;

X₆ is Leu or Thr;

X₇ is Lys or Thr;

X₈ is Leu or Lys;

X₉ is Met or Ser;

X₁₀ is Thr or Leu;

X₁₁ is Pro or Thr;

X₁₂ is Arg or Pro;

or combinations of such selections;

or

(g) said polypeptide includes an amino acid sequence of the following formula;

X₁-X₂-X₃-X₄-X₅-X₆-X₇-X₈-X₉-X₁₀-X₁₁-X₁₂-X₁₃ (SEQ ID NO;

7), and the following amino acid positions are independently selected as follows;

X₁ is Glu or Lys;

X₂ is His or Pro;

X₃ is Tyr;

X₄ is Asp or Gln;

X₅ is Asn or Thr;

X₆ is Asp or Pro;

X₇ is Ile or Pro;

X₈ is Leu or Thr;

X₉ is Lys;

X₁₀ is Gly or Met;

X₁₁ is Ala or Thr;

X₁₂ is Arg or His;

X₁₃ is His;

or combinations of such selections.

5. The BLyS binding polypeptide according to claim 3, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:

20–

162 as depicted in Tables 1–

8.

6. The BLyS binding polypeptide according to claim 3, comprising an amino acid sequence selected from the group consisting of:

AGKEPCYFYWECAVSGPGPEGGGK (SEQ ID NO;

163),

AGVPFCDLLTKHCFEAGPGPEGGGK (SEQ ID NO;

164),

GSSRLCHMDELHVCVHFAPPGPEGGGK (SEQ ID NO;

165),

GDGGNCYTDSLTKLHFCMGDEPGPEGGGK (SEQ ID NO;

166),

GYDVLTKLYFVPGGPGPEGGGK (SEQ ID NO;

167), and

WTDSLTGLWFPDGGPGPEGGGK (SEQ ID NO;

168).

7. A method for detecting BLyS or a BLyS-like polypeptide in a solution suspected of containing it comprising:

(a) contacting said solution with a polypeptide according to any of claim 1 or 3, and (b) determining whether binding has occurred between said polypeptide and BLyS or a BLyS-like polypeptide.

8. A method for purifying BLyS or a BLyS-like polypeptide comprising:

contacting a solution containing BLyS or a BLyS-like polypeptide to a support that comprises, immobilized thereon, a BLyS polypeptide according to claim 1 or 3; and

,

separating the solution from said support.

9. BLyS separation media comprising:

(a) a chromatographic matrix material, and, immobilized thereon,

(b) a BLyS binding molecule comprising a BLyS binding polypeptide as defined claim 1 or 3.

10. The BLyS separation media according to claim 9, comprising:

(a) a chromatographic matrix material, and, immobilized thereon,

(b) a BLyS binding molecule comprising a BLyS binding polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs;

20–

162 and 186–

435, as depicted in Tables 1–

8 and 14.

11. A method for separating BLyS or a BLyS-like polypeptide from a solution containing it comprising:

(a) contacting said solution with separation media as defined in claim 9,

(b) removing unbound material; and

(c) eluting bound BLyS or BLyS-like polypeptide from said separation media.

12. The polypeptide according to claim 1, wherein the polypeptide comprises an amino acid sequence according to H.

13. The polypeptide according to claim 12, wherein the polypeptide comprises

X₁-X₂-X₃-Cys-X₅-Phe-X₇-Trp-Glu-Cys-X₁₁-X₁₂-X₁₃ (SEQ ID NO:

1),

wherein

X₁ is Ala, Asn, Lys, or Ser;

X₂ is Ala, Glu, Met, Ser, or Val;

X₃ is Ala, Asn, Lys, or Pro;

X₁₁ is Ala, Gln, His, Phe, or Val;

X₁₂ is Asn, Gln, Gly, His, Ser, or Val; and

X₁₃ is Ala, Asn, Gly, Ile, Pro, or Ser.

14. The polypeptide according to claim 13, wherein X₁ is Lys.

15. The polypeptide according to claim 12, wherein X₅ is Tyr.

16. The polypeptide according to claim 12, wherein X₇ is Tyr.

17. The polypeptide according to claim 12, wherein X₅ is Tyr;

and X₇ is Tyr.

18. The polypeptide according to claim 12, that comprises SEQ ID NO:

22, 23, 24, 25, or 26.

19. The polypeptide according to claim 12, that comprises SEQ ID NO:

27.

20. The BLyS binding polypeptide according to claim 12, wherein the polypeptide comprises the sequence AGKEPCYFYWECAVSGPGPEGGGK (SEQ ID NO:

163).

21. The BLyS binding polypeptide of claim 1, wherein the polypeptide binds BLyS with an affinity less than 3 μ

M.

22. The BLyS binding polypeptide of claim 12, wherein the polypeptide binds BLyS with an affinity less than 3 μ

M.

23. The BLyS binding polypeptide of claim 13, wherein the polypeptide binds BLyS with an affinity less than 3 μ

M.

24. The BLyS binding polypeptide of claim 1, wherein the polypeptide binds BLyS at least 12-fold better than the polypeptide binds strepavidin.

25. The BLyS binding polypeptide of claim 12, wherein the polypeptide binds BLyS at least 12-fold better than the polypeptide binds strepavidin.

26. The BLyS binding polypeptide of claim 1, that comprises an amino acid sequence according to formula I.

27. The BLyS binding polypeptide of claim 26, that comprises SEQ ID NO:

28.

28. A method for purifying BLyS or a BLyS-like polypeptide, the method comprising:

contacting a solution containing BLyS or a BLyS-like polypeptide to a support that comprises, immobilized thereon, a BlyS binding polypeptide according to claim 12, 13, 14, 15, 16, 17, 18, 19, or 20; and

, separating the solution from the support.

29. The polypeptide according to claim 1, wherein the polypeptide comprises an amino acid sequence according to formula J.

30. The polypeptide according to claim 1, wherein the polypeptide comprises an amino acid sequence according to formula K.

31. The polypeptide according to claim 1, wherein the polypeptide comprises an amino acid sequence according to formula L.