BoNT/A peptides and methods of predicting and reducing immunoresistance to botulinum toxin therapy
View Patent ↗The present invention provides BoNT/A peptides as well as methods of predicting or determining immunoresistance to botulinum toxin therapy in an individual using BoNT/A peptides.
1. A therapeutic immunoadsorption method of removing an anti-botulinum toxin antibody from an individual in need thereof, the method comprising the steps of:
(a) removing blood from said individual;
(b) contacting said blood, or an antibody-containing component thereof, with a BoNT/A peptide immunoreactive with an anti-botulinum toxin antibody under conditions suitable for forming an immunoreactive complex between said immunoreactive BoNT/A peptide and said anti-botulinum toxin antibody;
wherein said immunoreactive BoNT/A peptide has a length of at most 60 amino acids, said BoNT/A peptide comprising amino acids 785-803 of SEQ ID NO: 1, a conservative BoNT/A amino acid sequence variant of amino acids 785-803 of SEQ ID NO: 1, or an immunoreactive BoNT/A amino acid sequence fragment of amino acids 785-803 of SEQ ID NO: 1;
wherein said antibodies immunoreact with amino acids 785-803 of SEQ ID NO: 1, a conservative BoNT/A amino acid sequence variant of amino acids 785-803 of SEQ ID NO: 1, or an immunoreactive BoNT/A amino acid sequence fragment of amino acids 785-803 of SEQ ID NO: 1:
wherein said conservative BoNT/A amino acid sequence variant immunoreactive with said anti-botulinum toxin antibody comprises 1-4 conservative amino acid substitutions to amino acids 785-803 of SEQ ID NO: 1; and
wherein said immunoreactive BoNT/A amino acid sequence fragment immunoreactive with said anti-botulinum toxin antibody comprises at least six consecutive amino acids of 785-803 of SEQ ID NO: 1;
(c) removing said complex from said blood, or antibody-containing component thereof, thereby forming anti-botulinum toxin antibody-depleted blood, or an anti-botulinum toxin antibody-depleted component thereof; and
(d) returning said anti-botulinum toxin antibody-depleted blood, or said anti-botulinum toxin antibody-depleted component thereof, to said individual.
2. The therapeutic immunoadsorption method of claim 1 , wherein said BoNT/A peptide comprises amino acids 785-803 of SEQ ID NO: 1.
3. The therapeutic immunoadsorption method of claim 1 , wherein said immunoreactive BoNT/A peptide is attached to a solid support.
4. The therapeutic immunoadsorption method of claim 3 , wherein said immunoreactive BoNT/A peptide is covalently bound to said solid support or noncovalently bound to said solid support.
5. The therapeutic immunoadsorption method of claim 1 , wherein said immunoreactive BoNT/A peptide has a length of at most 40 amino acids.
6. The therapeutic immunoadsorption method of claim 1 , wherein said immunoreactive BoNT/A peptide has a length of at most 25 amino acids.
7. A therapeutic immunoadsorption method of removing an anti-botuinum toxin antibody from an individual in need thereof, the method comprising the steps of:
(a) removing blood from said individual;
(b) contacting said blood, or an antibody-containing component thereof, with at least one BoNT/A peptide immunoreactive with an anti-botulinum toxin antibody under conditions suitable for forming an immunoreactive complex between at least one of said immunoreactive BoNT/A peptide and said anti-botulinum toxin antibody;
wherein one of said immunoreactive BoNT/A peptides has a length of at most 60 amino acids, said BoNT/A peptide comprising amino acids 785-803 of SEQ ID NO: 1, a conservative BoNT/A amino acid sequence variant of amino acids 785-803 of SEQ ID NO: 1, or an immunoreactive BoNT/A amino acid sequence fragment of amino acids 785-803 of SEQ ID NO: 1;
wherein said antibodies immunoreact with amino acids 785-803 of SEQ ID NO: 1, a conservative BoNT/A amino acid sequence variant of amino acids 785-803 of SEQ ID NO: 1, or an immunoreactive BoNT/A amino acid sequence fragment of amino acids 785-803 of SEQ ID NO: 1:
wherein said conservative BoNT/A amino acid sequence variant immunoreactive with said anti-botulinum toxin antibody comprises 1-4 conservative amino acid substitutions to amino acids 785-803 of SEQ ID NO: 1; and
wherein said immunoreactive BoNT/A amino acid sequence fragment immunoreactive with said anti-botulinum toxin antibody comprises at least six consecutive amino acids of 785-803 of SEQ ID NO: 1;
(c) removing said complex from said blood, or antibody-containing component thereof, thereby forming anti-botulinum toxin antibody-depleted blood, or an anti-botulinum toxin antibody-depleted component thereof; and
(d) returning said anti-botulinum toxin antibody-depleted blood, or said anti-botulinum toxin antibody-depleted component thereof, to said individual.
8. The therapeutic immunoadsorption method of claim 7 , wherein in said step (b), said blood, or an antibody-containing component thereof, is contacted with two or more immunoreactive BoNT/A peptides.
9. The therapeutic immunoadsorption method of claim 8 , wherein in said step (b), said blood, or an antibody-containing component thereof, is contacted with two immunoreactive BoNT/A peptides.
10. The therapeutic immunoadsorption method of claim 8 , wherein in said step (b), said blood, or an antibody-containing component thereof, is contacted with three immunoreactive BoNT/A peptides.
11. The therapeutic immunoadsorption method of claim 7 , wherein each of said immunoreactive BoNT/A peptides is attached to a solid support.
12. The therapeutic immunoadsorption method of claim 11 , wherein each of said attached immunoreactive BoNT/A peptides is covalently bound to said solid support or noncovalently bound to said solid support.
13. The therapeutic immunoadsorption method of claim 7 , wherein each of said immunoreactive BoNT/A peptides has a length of at most 40 amino acids.
14. The therapeutic immunoadsorption method of claim 7 , wherein each of said immunoreactive BoNT/A peptides has a length of at most 25 amino acids.
15. The therapeutic immunoadsorption method of either claim 1 or claim 7 , wherein said antibody-containing component is plasma or serum.
16. The therapeutic immunoadsorption method of either claim 1 or claim 7 , wherein said anti-botulinum toxin antibody is an anti-botulinum toxin type A antibody.
17. The therapeutic immunoadsorption method of either claim 1 or claim 7 , further comprising the step of pre-clearing a class of antibody from said blood, or said antibody-containing component thereof, prior to performing said step (b).
18. The therapeutic immunoadsorption method of claim 17 , wherein said class of antibody is an lgG class.
19. The therapeutic immunoadsorption method of either claim 1 or claim 7 , further comprising the step of assaying said antibody-depleted blood, or antibody-depleted component thereof, for the presence or absence of anti-botulinum toxin antibodies prior to performing said step (d).