Isolation of therapeutic target specific VNAR domains to ICOSL
The present invention provides ICOSL specific antigen binding molecules which are isolated from immunized and synthetic Elasmobranchii derived libraries. In particular, the present invention relates to shark Variable New Antigen Receptor (VNAR) domains that specifically bind and neutralize the activity of human Induced Co-Stimulatory Ligand (ICOSL). The neutralizing VNAR domains are isolated from two independent sources; an immunized nurse shark library and a synthetic spiny dogfish framework fusion library. The molecules may be formulated as pharmaceutical compositions and used in medicine.
1. An ICOSL specific antigen binding molecule comprising an amino acid sequence represented by the formula (I)
A-X-B-Y-C (I);
wherein:
A is SEQ ID NO: 1, SEQ ID NO: 4 or SEQ ID NO: 7, or a sequence at least 87% identical to SEQ ID NO: 1, SEQ ID NO: 4 or SEQ ID NO: 7,
X is a CDR1 region of SEQ ID NO: 95,
B is SEQ ID NO: 2, SEQ ID NO: 5 or SEQ ID NO: 8, or a sequence at least 87% identical to SEQ ID NO: 2, SEQ ID NO: 5 or SEQ ID NO: 8,
Y is a CDR3 region of SEQ ID NO: 122, and
C is SEQ ID NO: 3, SEQ ID NO: 6, or SEQ ID NO: 9, or a sequence at least 87% identical to SEQ ID NO: 3, SEQ ID NO: 6, or SEQ ID NO: 9,
in which:
SEQ ID NO: 1 is
TRVDQTPRTATKETGESLTINCVLTDT,
TRVDQTPRTATKETGESLTINCWTGA
SEQ ID NO: 2 is
TSWFRKNPGTTDWERMSIGGRYVESVNKGAKSFSLRIKDLTVADSATYY
CKA
or
TSWFRKNPGTTDWERMSIGGRYVESVNKGAKSFSLRIKDLTVADSATYI
CRA
SEQ ID NO: 3 is
DGAGTVLTVN
SEQ ID NO: 4 is
ASVNQTPRTATKETGESLTINCVLTDT
SEQ ID NO: 5 is
TYWYRKNPGSSNQERISISGRYVESVNKRTMSFSLRIKDLTVADSATYY
CKA
or
TYWYRKNPGSSNQERISISGRYVESVNKRTMSFSLRIKDLTVADSATYI
CRA
SEQ ID NO: 6 is
YGAGTVLTVN
SEQ ID NO: 7 is
ARVDQTPRSVTKETGESLTINCVLRDP
or
ARVDQTPRSVTKETGESLTINCVLRDA
or
ARVDQTPRSVTKETGESLTINCVLRDG
or
ARVDQTPRSVTKETGESLTINCVLRES
SEQ ID NO: 8 is
TCWYRKKSGSTNEESISKGGRYVETVNSGSKSFSLRINDLTVEDGGTYR
CGA
or
TCWSRKKSGSTNEESISKGGRYVETVNSGSKSFSLRINDLTVEDGGTYR
CGL,
TCWTRKKSGSTNEESISKGGRYVETVNSGSKSFSLRINDLTVEDGGTYR
CAL,
TCWYRKKSGSTNEESISKGGRYVETVNSGSKSFSLRINDLTVEDGGTYR
CGV,
TCWYRKKSGSTNEESISKGGRYVETVNSGSKSFSLRINDLTVEDGGTYR
CGH,
TCWYRKKSGSTNEESISKGGRYVETVNSGSKSFSLRISDLTVEDGGTYR
CGH,
SEQ ID NO: 9 is
CGGGTVVTVN, CGGGTAVTVN, CGDGTAVTVN,
or
CGDGTAVTVN,
SEQ ID NO: 95 is DYGLFS, and
SEQ ID NO: 122 is FTWPWEWPDRWFRPWY.
2. The ICOSL specific antigen binding molecule as claimed in claim 1 which is humanized.
3. A fusion protein comprising the ICOSL specific antigen binding molecule as claimed in claim 1 .
4. The fusion protein as claimed in claim 3 , in which the ICOSL specific antigen binding molecule is fused to a biologically active protein.
5. A pharmaceutical composition comprising the ICOSL specific antigen binding molecule as defined in claim 1 .
6. The ICOSL specific antigen binding molecule of claim 1 , wherein the ICOSL specific antigen binding molecule comprises the amino acid sequence of SEQ ID NO: 21.