Agrochemical compositions comprising antibodies binding to sphingolipids

Abstract

This disclosure relates to agrochemical and biological control compositions for combating pests, more specifically plant pests, comprising at least one polypeptide, which specifically binds to a pest. The disclosure further provides methods for protecting or treating a plant or a part of a plant from an infection or other biological interaction with a plant pathogen, at least comprising the step of applying directly or indirectly to a plant or to a part of a plant, an agrochemical composition, under conditions effective to protect or treat a plant or a part of a plant against a infection or biological interaction with a plant pathogen. Further provided are methods for producing such agrochemical compositions and formulations, to polypeptides with a specific pesticidal activity comprised within an agrochemical formulation, to nucleic acids encoding such polypeptide and to plants comprising chimeric genes comprising such nucleic acids.

1 - 15 . (canceled) 
     
     
         16 . A polypeptide which comprises: (a) one or more of the combinations chosen from the group consisting of:
 a CDR1 region having SEQ ID NO: 85, a CDR2 region having has SEQ ID NO: 169, and a CDR3 region having SEQ ID NO: 253, and/or   a CDR1 region having SEQ ID NO: 86, a CDR2 region having has SEQ ID NO: 170, and a CDR3 region having SEQ ID NO: 254, and/or   a CDR1 region having SEQ ID NO: 87, a CDR2 region having has SEQ ID NO: 171, and a CDR3 region having SEQ ID NO: 255, and/or   a CDR1 region having SEQ ID NO: 88, a CDR2 region having has SEQ ID NO: 172, and a CDR3 region having SEQ ID NO: 256, and/or   a CDR1 region having SEQ ID NO: 89, a CDR2 region having has SEQ ID NO: 173, and a CDR3 region having SEQ ID NO: 257, and/or   a CDR1 region having SEQ ID NO: 90, a CDR2 region having has SEQ ID NO: 174, and a CDR3 region having SEQ ID NO: 258, and/or   a CDR1 region having SEQ ID NO: 91, a CDR2 region having has SEQ ID NO: 175, and a CDR3 region having SEQ ID NO: 259, and/or   a CDR1 region having SEQ ID NO: 92, a CDR2 region having has SEQ ID NO: 176, and a CDR3 region having SEQ ID NO: 260, and/or   a CDR1 region having SEQ ID NO: 93, a CDR2 region having has SEQ ID NO: 177, and a CDR3 region having SEQ ID NO: 261, and/or   a CDR1 region having SEQ ID NO: 94, a CDR2 region having has SEQ ID NO: 178, and a CDR3 region having SEQ ID NO: 262, and/or   a CDR1 region having SEQ ID NO: 95, a CDR2 region having has SEQ ID NO: 179, and a CDR3 region having SEQ ID NO: 263, and/or   a CDR1 region having SEQ ID NO: 96, a CDR2 region having has SEQ ID NO: 180, and a CDR3 region having SEQ ID NO: 264, and/or   a CDR1 region having SEQ ID NO: 97, a CDR2 region having has SEQ ID NO: 181, and a CDR3 region having SEQ ID NO: 265, and/or   a CDR1 region having SEQ ID NO: 98, a CDR2 region having has SEQ ID NO: 182, and a CDR3 region having SEQ ID NO: 266, and/or   a CDR1 region having SEQ ID NO: 99, a CDR2 region having has SEQ ID NO: 183, and a CDR3 region having SEQ ID NO: 267, and/or   a CDR1 region having SEQ ID NO: 100, a CDR2 region having has SEQ ID NO: 184, and a CDR3 region having SEQ ID NO: 268, and/or   a CDR1 region having SEQ ID NO: 101, a CDR2 region having has SEQ ID NO: 185, and a CDR3 region having SEQ ID NO: 269, and/or   a CDR1 region having SEQ ID NO: 102, a CDR2 region having has SEQ ID NO: 186, and a CDR3 region having SEQ ID NO: 270, and/or   a CDR1 region having SEQ ID NO: 103, a CDR2 region having has SEQ ID NO: 187, and a CDR3 region having SEQ ID NO: 271, and/or   a CDR1 region having SEQ ID NO: 104, a CDR2 region having has SEQ ID NO: 188, and a CDR3 region having SEQ ID NO: 272, and/or   a CDR1 region having SEQ ID NO: 105, a CDR2 region having has SEQ ID NO: 189, and a CDR3 region having SEQ ID NO: 273, and/or   a CDR1 region having SEQ ID NO: 106, a CDR2 region having has SEQ ID NO: 190, and a CDR3 region having SEQ ID NO: 274, and/or   a CDR1 region having SEQ ID NO: 107, a CDR2 region having has SEQ ID NO: 191, and a CDR3 region having SEQ ID NO: 275, and/or   a CDR1 region having SEQ ID NO: 108, a CDR2 region having has SEQ ID NO: 192, and a CDR3 region having SEQ ID NO: 276, and/or   a CDR1 region having SEQ ID NO: 109, a CDR2 region having has SEQ ID NO: 193, and a CDR3 region having SEQ ID NO: 277, and/or   a CDR1 region having SEQ ID NO: 110, a CDR2 region having has SEQ ID NO: 194, and a CDR3 region having SEQ ID NO: 278, and/or   a CDR1 region having SEQ ID NO: 111, a CDR2 region having has SEQ ID NO: 195, and a CDR3 region having SEQ ID NO: 279, and/or   a CDR1 region having SEQ ID NO: 112, a CDR2 region having has SEQ ID NO: 196, and a CDR3 region having SEQ ID NO: 280, and/or   a CDR1 region having SEQ ID NO: 113, a CDR2 region having has SEQ ID NO: 197, and a CDR3 region having SEQ ID NO: 281, and/or   a CDR1 region having SEQ ID NO: 114, a CDR2 region having has SEQ ID NO: 198, and a CDR3 region having SEQ ID NO: 282, and/or   a CDR1 region having SEQ ID NO: 115, a CDR2 region having has SEQ ID NO: 199, and a CDR3 region having SEQ ID NO: 283, and/or   a CDR1 region having SEQ ID NO: 116, a CDR2 region having has SEQ ID NO: 200, and a CDR3 region having SEQ ID NO: 284, and/or   a CDR1 region having SEQ ID NO: 117, a CDR2 region having has SEQ ID NO: 201, and a CDR3 region having SEQ ID NO: 285, and/or   a CDR1 region having SEQ ID NO: 118, a CDR2 region having has SEQ ID NO: 202, and a CDR3 region having SEQ ID NO: 286, and/or   a CDR1 region having SEQ ID NO: 119, a CDR2 region having has SEQ ID NO: 203, and a CDR3 region having SEQ ID NO: 287, and/or   a CDR1 region having SEQ ID NO: 120, a CDR2 region having has SEQ ID NO: 204, and a CDR3 region having SEQ ID NO: 288, and/or   a CDR1 region having SEQ ID NO: 121, a CDR2 region having has SEQ ID NO: 205, and a CDR3 region having SEQ ID NO: 289, and/or   a CDR1 region having SEQ ID NO: 122, a CDR2 region having has SEQ ID NO: 206, and a CDR3 region having SEQ ID NO: 290, and/or   a CDR1 region having SEQ ID NO: 123, a CDR2 region having has SEQ ID NO: 207, and a CDR3 region having SEQ ID NO: 291, and/or   a CDR1 region having SEQ ID NO: 124, a CDR2 region having has SEQ ID NO: 208, and a CDR3 region having SEQ ID NO: 292, and/or   a CDR1 region having SEQ ID NO: 125, a CDR2 region having has SEQ ID NO: 209, and a CDR3 region having SEQ ID NO: 293, and/or   a CDR1 region having SEQ ID NO: 126, a CDR2 region having has SEQ ID NO: 210, and a CDR3 region having SEQ ID NO: 294, and/or   a CDR1 region having SEQ ID NO: 127, a CDR2 region having has SEQ ID NO: 211, and a CDR3 region having SEQ ID NO: 295, and/or   a CDR1 region having SEQ ID NO: 128, a CDR2 region having has SEQ ID NO: 212, and a CDR3 region having SEQ ID NO: 296, and/or   a CDR1 region having SEQ ID NO: 129, a CDR2 region having has SEQ ID NO: 213, and a CDR3 region having SEQ ID NO: 297, and/or   a CDR1 region having SEQ ID NO: 130, a CDR2 region having has SEQ ID NO: 214, and a CDR3 region having SEQ ID NO: 298, and/or   a CDR1 region having SEQ ID NO: 131, a CDR2 region having has SEQ ID NO: 215, and a CDR3 region having SEQ ID NO: 299, and/or   a CDR1 region having SEQ ID NO: 132, a CDR2 region having has SEQ ID NO: 216, and a CDR3 region having SEQ ID NO: 300, and/or   a CDR1 region having SEQ ID NO: 133, a CDR2 region having has SEQ ID NO: 217, and a CDR3 region having SEQ ID NO: 301, and/or   a CDR1 region having SEQ ID NO: 134, a CDR2 region having has SEQ ID NO: 218, and a CDR3 region having SEQ ID NO: 302, and/or   a CDR1 region having SEQ ID NO: 135, a CDR2 region having has SEQ ID NO: 219, and a CDR3 region having SEQ ID NO: 303, and/or   a CDR1 region having SEQ ID NO: 136, a CDR2 region having has SEQ ID NO: 220, and a CDR3 region having SEQ ID NO: 304, and/or   a CDR1 region having SEQ ID NO: 137, a CDR2 region having has SEQ ID NO: 221, and a CDR3 region having SEQ ID NO: 305, and/or   a CDR1 region having SEQ ID NO: 138, a CDR2 region having has SEQ ID NO: 222, and a CDR3 region having the amino acid sequence NRY, and/or   a CDR1 region having SEQ ID NO: 139, a CDR2 region having has SEQ ID NO: 223, and a CDR3 region having SEQ ID NO: 306, and/or   a CDR1 region having SEQ ID NO: 140, a CDR2 region having has SEQ ID NO: 224, and a CDR3 region having SEQ ID NO: 307, and/or   a CDR1 region having SEQ ID NO: 141, a CDR2 region having has SEQ ID NO: 225, and a CDR3 region having SEQ ID NO: 308, and/or   a CDR1 region having SEQ ID NO: 142, a CDR2 region having has SEQ ID NO: 226, and a CDR3 region having SEQ ID NO: 309, and/or   a CDR1 region having SEQ ID NO: 143, a CDR2 region having has SEQ ID NO: 227, and a CDR3 region having SEQ ID NO: 310, and/or   a CDR1 region having SEQ ID NO: 144, a CDR2 region having has SEQ ID NO: 228, and a CDR3 region having SEQ ID NO: 311, and/or   a CDR1 region having SEQ ID NO: 145, a CDR2 region having has SEQ ID NO: 229, and a CDR3 region having SEQ ID NO: 312, and/or   a CDR1 region having SEQ ID NO: 146, a CDR2 region having has SEQ ID NO: 230, and a CDR3 region having SEQ ID NO: 313, and/or   a CDR1 region having SEQ ID NO: 147, a CDR2 region having has SEQ ID NO: 231, and a CDR3 region having SEQ ID NO: 314, and/or   a CDR1 region having SEQ ID NO: 148, a CDR2 region having has SEQ ID NO: 232, and a CDR3 region having SEQ ID NO: 315, and/or   a CDR1 region having SEQ ID NO: 149, a CDR2 region having has SEQ ID NO: 233, and a CDR3 region having SEQ ID NO: 316, and/or   a CDR1 region having SEQ ID NO: 150, a CDR2 region having has SEQ ID NO: 234, and a CDR3 region having SEQ ID NO: 317, and/or   a CDR1 region having SEQ ID NO: 151, a CDR2 region having has SEQ ID NO: 235, and a CDR3 region having SEQ ID NO: 318, and/or   a CDR1 region having SEQ ID NO: 152, a CDR2 region having has SEQ ID NO: 236, and a CDR3 region having SEQ ID NO: 319, and/or   a CDR1 region having SEQ ID NO: 153, a CDR2 region having has SEQ ID NO: 237, and a CDR3 region having SEQ ID NO: 320, and/or   a CDR1 region having SEQ ID NO: 154, a CDR2 region having has SEQ ID NO: 238, and a CDR3 region having SEQ ID NO: 321, and/or   a CDR1 region having SEQ ID NO: 155, a CDR2 region having has SEQ ID NO: 239, and a CDR3 region having SEQ ID NO: 322, and/or   a CDR1 region having SEQ ID NO: 156, a CDR2 region having has SEQ ID NO: 240, and a CDR3 region having SEQ ID NO: 323, and/or   a CDR1 region having SEQ ID NO: 157, a CDR2 region having has SEQ ID NO: 241, and a CDR3 region having SEQ ID NO: 324, and/or   a CDR1 region having SEQ ID NO: 158, a CDR2 region having has SEQ ID NO: 242, and a CDR3 region having SEQ ID NO: 325, and/or   a CDR1 region having SEQ ID NO: 159, a CDR2 region having has SEQ ID NO: 243, and a CDR3 region having SEQ ID NO: 326, and/or   a CDR1 region having SEQ ID NO: 160, a CDR2 region having has SEQ ID NO: 244, and a CDR3 region having SEQ ID NO: 327, and/or   a CDR1 region having SEQ ID NO: 161, a CDR2 region having has SEQ ID NO: 245, and a CDR3 region having SEQ ID NO: 328, and/or   a CDR1 region having SEQ ID NO: 162, a CDR2 region having has SEQ ID NO: 246, and a CDR3 region having SEQ ID NO: 329, and/or   a CDR1 region having SEQ ID NO: 163, a CDR2 region having has SEQ ID NO: 247, and a CDR3 region having SEQ ID NO: 330, and/or   a CDR1 region having SEQ ID NO: 164, a CDR2 region having has SEQ ID NO: 248, and a CDR3 region having SEQ ID NO: 331, and/or   a CDR1 region having SEQ ID NO: 165, a CDR2 region having has SEQ ID NO: 249, and a CDR3 region having SEQ ID NO: 332, and/or   a CDR1 region having SEQ ID NO: 166, a CDR2 region having has SEQ ID NO: 250, and a CDR3 region having SEQ ID NO: 333, and/or   a CDR1 region having SEQ ID NO: 167, a CDR2 region having has SEQ ID NO: 251, and a CDR3 region having SEQ ID NO: 334, and/or   a CDR1 region having SEQ ID NO: 168, a CDR2 region having has SEQ ID NO: 252, and a CDR3 region having SEQ ID NO: 335; or   (b) a polypeptide comprising a sequence having at least 85% sequence identity to any one of the polypeptides of (a).   
     
     
         17 . A method for the preparation of a heavy chain variable domain of a heavy chain antibody (VHH) which specifically binds to and/or has affinity to a plant pest target, which method comprises:
 a) providing a set, collection or library of random amino acid sequences;   b) screening said set, collection or library of amino acid sequences that can bind to and/or have affinity for the pest target; and   c) isolating the amino acid sequence(s) that can bind to and/or have affinity for the pest target,
 thereby to prepare a VHH which specifically binds to and/or has affinity to a plant pest target. 
   
     
     
         18 . A method for the preparation of a heavy chain variable domain of a heavy chain antibody (VHH) according to  claim 17 , wherein the set, collection or library of polypeptide sequences in a) is selected from the group consisting of: a set, collection or library of immunoglobulin fragment sequences, such as a naïve set, collection or library of immunoglobulin fragment sequences; a synthetic or semi-synthetic set, collection or library of immunoglobulin fragment sequences; and a set, collection or library of immunoglobulin fragment sequences that have been subjected to affinity maturation. 
     
     
         19 . A method according to  claim 17 , wherein the set, collection or library of polypeptide sequences in b) may be displayed on a phage, phagemid, ribosome or suitable micro-organism, such as a yeast. 
     
     
         20 . A method according to  claim 17 , wherein prior to step (a) the method comprises: immunizing an animal with a pest target or with a suitable antigenic determinant based thereon or derived therefrom, such as an antigenic part, fragment, region, domain, loop or other epitope thereof, optionally wherein said animal is a camelid or a shark, preferably wherein the camelid is a llama. 
     
     
         21 . A method according to  claim 20 , wherein the antigenic determinant is an extracellular part, region, domain, loop or other extracellular epitope(s). 
     
     
         22 . A method for the preparation of a heavy chain variable domain of a heavy chain antibody (VHH) which specifically binds to and/or has affinity to a plant pest target, which method comprises:
 a) providing a collection or sample of cells expressing polypeptide sequences;   b) screening said collection or sample of cells for cells that express an amino acid sequence that binds to and/or has affinity for the plant pest target; and   c) either (i) isolating said amino acid sequence; or (ii) isolating from said cell a nucleic acid sequence that encodes said amino acid sequence, followed by expressing said amino acid sequence,
 thereby to prepare a VHH which specifically binds to and/or has affinity to a plant pest target. 
   
     
     
         23 . A method according to  claim 22 , wherein the collection or sample of cells in a) is a collection or sample of B-cells. 
     
     
         24 . A method according to  claim 22 , wherein prior to step (a) the method comprises: immunizing an animal with a pest target or with a suitable antigenic determinant based thereon or derived therefrom, such as an antigenic part, fragment, region, domain, loop or other epitope thereof. 
     
     
         25 . A method according to  claim 24 , wherein said animal is a camelid or a shark, preferably wherein the camelid is a llama. 
     
     
         26 . A method for the preparation of a heavy chain variable domain of a heavy chain antibody (VHH) which specifically binds to and/or has affinity to a plant pest target, which method comprises:
 a) providing a set, collection or library of nucleic acid sequences encoding a polypeptide amino acid sequence;   b) screening said set, collection or library of nucleic acid sequences for nucleic acid sequences that encode an amino acid sequence that binds to and/or has affinity for the pest target; and   c) isolating said nucleic acid sequence, followed by expressing said amino acid sequence,
 thereby to prepare a VHH which specifically binds to and/or has affinity to a plant pest target. 
   
     
     
         27 . A method according to  claim 26 , wherein the set, collection or library of nucleic acid sequences encoding amino acid sequences in a) is selected from the group consisting of: a set, collection or library of nucleic acid sequences encoding a naïve set, collection or library of immunoglobulin fragment sequences; a set, collection or library of nucleic acid sequences encoding a synthetic or semi-synthetic set, collection or library of immunoglobulin fragment sequences; and a set, collection or library of nucleic acid sequences encoding a set, collection or library of immunoglobulin fragment sequences that have been subjected to affinity maturation. 
     
     
         28 . A method according to  claim 26 , wherein the set, collection or library of nucleic acid sequences in a) encodes a set, collection or library of such as VH domains or VHH domains. 
     
     
         29 . A method according to  claim 26 , wherein the set, collection or library of nucleic acid sequences in a) may be displayed on a phage, phagemid, ribosome or suitable micro-organism, such as a yeast. 
     
     
         30 . A method according to  claim 26  wherein prior to step (a) the method comprises: immunizing an animal with a pest target or with a suitable antigenic determinant based thereon or derived therefrom, such as an antigenic part, fragment, region, domain, loop or other epitope thereof; optionally wherein said animal is a camelid or shark, preferably wherein the camelid is a llama. 
     
     
         31 . A method for the preparation of an agrochemical composition which method comprises:
 a) obtaining a VHH which specifically binds to and/or has affinity to a plant pest target using a method according to  claim 17 ; and   b) formulating the VHH or functional fragment thereof in an agrochemical composition, thereby to prepare an agrochemical composition.   
     
     
         32 . A method according to  claim 17 , wherein said VHH which specifically binds to and/or has affinity to a plant pest target specifically binds to at least one plant pathogenic fungus. 
     
     
         33 . A method according to  claim 17 , wherein said VHH which specifically binds to and/or has affinity to a plant pest target specifically binds to at least one plasma membrane component of said plant pest target. 
     
     
         34 . A method according to  claim 33 , wherein said at least one plasma membrane component of said pest is a sphingolipid. 
     
     
         35 . A method according to  claim 34 , wherein said at least one plasma membrane component of said pest is glucosylceramide.