1. A method of preparing a recombinant allergen derived from a naturally occurring allergen, wherein specific IgE binding to the recombinant allergen is reduced compared to the IgE binding to said naturally occurring allergen, which method comprises:
a) identifying amino acid residues in said naturally occurring allergen which are conserved with more than 70% identity in all of the known homologous proteins within the taxonomic order from which said naturally occurring allergen originates;
b) defining at least one patch of said conserved amino acid residues which patch (i) is connected over at least 400 2 of the surface of the three-dimensional structure of said naturally occurring allergen molecule; and (ii) comprises at least one B cell epitope, and wherein (iii) said conserved amino acid residues have a solvent accessibility of at least 20%; and
c) substituting at least one of said identified amino acid residues in said defined at least one patch with a non-conservative amino acid residue,
wherein the -carbon backbone tertiary structure of the recombinant allergen is essentially preserved as compared with the -carbon backbone tertiary structure of said naturally occurring allergen.
2. The method of claim 1, wherein the amino acid residues identified in step (a) are 90% or more conserved.
3. The method of claim 1, which method further comprises:
b) ranking said conserved amino acid residues within said at least one patch in order of percent solvent accessibility; and
b) selecting said at least one amino acid residue for substitution in step (c) from the amino acid residues having a solvent accessibility of 20-80%.
4. The method of claim 1, wherein the percent solvent accessibility is in a range from 30-80%.
5. The method of claim 1 in which the average root mean square of the atomic coordinates of the -carbon backbone tertiary structure of the recombinant allergen deviates from the average root mean square of the atomic coordinates of the -carbon backbone tertiary structure of said naturally occurring allergen by less than 2 .
6. The method of claim 1, wherein the amino acid residue substitution in step (c) is performed using site-directed mutagenesis.
7. The method of claim 1, wherein said at least one amino acid residue being substituted is a polar amino acid and said non-conservative amino acid is a polar amino acid.
8. The method of claim 1, wherein said at least one amino acid residue being substituted is a non-polar amino acid and said non-conservative amino acid is a non-polar amino acid.
9. The method of claim 1, wherein said naturally occurring allergen is an inhalation allergen or venom allergen.
10. The method of claim 1, wherein said naturally occurring allergen is a tree pollen allergen, grass pollen allergen, weed pollen allergen, herb pollen allergen, fungal allergen, dust mite allergen, animal allergen or insect allergen.
11. The method of claim 10 wherein
(a) said tree pollen allergen is from a tree taxonomically belonging to Fagales, Oleales, or Pineales;
(b) said grass pollen allergen is from a grass taxonomically belonging to Poales;
(c) said herb pollen allergen is from an herb taxonomically belonging to Aasterales;
(d) said fungal allergen is from a fungus taxonomically belonging to Alternaria or Cladosporium;
(e) said dust mite allergen is from a dust mite taxonomically belonging to Dermatophagoides;
(f) said animal allergen is from an animal taxonomically belonging to Felis, Canis, or Equus;
(g) said insect allergen is from an insect taxonomically belonging to Blatella, Periplaneta, or Hymenoptera;
(h) said weed pollen allergen is from a weed taxonomically belonging to Urticales.
12. The method of claim 11 wherein
(a) said tree taxonomically belonging to Fagales belongs taxonomically to Betula, Alnus, Corylus, or Carpinus;
(b) said tree taxonomically belonging to Oleales belongs taxonomically to Olea;
(c) said tree taxonomically belonging to Pineales belongs taxonomically to Cryptomeria;
(d) said grass taxonomically belonging to Poales belongs taxonomically to Cynodon, Dactylis, Festuca, Holcus, Lolium, Phleum, Poa, Secale or Sorghum;
(e) said herb taxonomically belonging to Aasterales belongs taxonomically to Ambrosia, Artemicia, or Urticales;
(f) said fungus taxonomically belonging to Alternaria belongs taxonomically to Alternata;
(g) said fungus taxonomically belonging to Cladosporium belongs taxonomically to herbarium;
(h) said dust mite belonging taxonomically belonging to Dermatophagoides belongs taxonomically to Farinae or Pteronyssinus;
(i) said animal taxonomically belonging to Felis belongs taxonomically to Domesticus;
(j) said animal taxonomically belonging to Canis belongs taxonomically to familiaris;
(k) said animal taxonomically belonging to Equus belongs taxonomically to caballus;
(l) said insect taxonomically belonging to Blatella belongs taxonomically to Germanica;
(m) said insect taxonomically belonging to Periplaneta belongs taxonomically to americana;
(n) said insect taxonomically belonging to Hymenoptera belongs taxonomically to Vespidae, Apidae; and
(o) said weed taxonomically belonging to Urticales belongs taxonomically to Parietaria.
13. A recombinant allergen derived from a naturally occurring allergen, wherein at least one solvent-accessible amino acid which is essentially conserved with more than 70% identity in all of the known homologous proteins within the taxonomic order from which said naturally occurring allergen originates, is substituted with an amino acid residue that is not conserved in homologous naturally-occurring allergens within said taxonomic order, wherein the -carbon backbone tertiary structure of the recombinant allergen is conserved as compared with said naturally occurring allergen, and wherein specific IgE binding to the mutant allergen is reduced compared to IgE binding to said naturally-occurring allergen.
14. The recombinant allergen of claim 13, wherein said at least one solvent-accessible amino acid residue has a solvent accessibility of 20-80%.
15. The recombinant allergen of claim 13, in which between 1 and 5 solvent-accessible amino acid residues per at least 400 2, are substituted.
16. The recombinant allergen of claim 13, wherein the average root mean square deviation of the atomic coordinates comparing the -carbon backbone tertiary structures of said recombinant and said naturally occurring allergen is less than 2 .
17. The recombinant allergen of claim 13, wherein the specific IgE binding to said recombinant allergen is reduced by at least 5-10%.
18. The recombinant allergen of claim 17, wherein the specific IgE binding to said recombinant allergen is reduced by at least 10%.
19. The recombinant allergen of claim 13, wherein said naturally occurring allergen is an inhalation allergen or venom allergen.
20. The recombinant allergen of claim 13, wherein said naturally occurring allergen is a tree pollen allergen, grass pollen allergen, weed pollen allergen, herb pollen allergen, fungal allergen, dust mite allergen, animal allergen or insect allergen.
21. The recombinant allergen of claim 20 wherein
(a) said tree pollen allergen is from a tree taxonomically belonging to Fagales, Oleales, or Pineales;
(b) said grass pollen allergen is from a grass taxonomically belonging to Poales;
(c) said herb pollen allergen is from an herb taxonomically belonging to Aasterales;
(d) said fungal allergen is from a fungus taxonomically belonging to Alternaria or Cladosporium;
(e) said dust mite allergen is from a dust mite taxonomically belonging to Dermatophagoides;
(f) said animal allergen is from an animal taxonomically belonging to Felis, Canis, or Equus;
(g) said insect allergen is from an insect taxonomically belonging to Blatella, Periplaneta, or Hymenoptera;
(h) said weed pollen allergen is from a weed taxonomically belonging to Urticales.
22. The recombinant allergen of claim 21 wherein
(a) said tree taxonomically belonging to Fagales belongs taxonomically to Betula, Alnus, Corylus, or Carpinus;
(b) said tree taxonomically belonging to Oleales belongs taxonomically to Olea;
(c) said tree taxonomically belonging to Pineales belongs taxonomically to Cryptomeria;
(d) said grass taxonomically belonging to Poales belongs taxonomically to Cynodon, Dactylis, Festuca, Holcus, Lolium, Phleum, Poa, Secale or Sorghum;
(e) said herb taxonomically belonging to Aasterales belongs taxonomically to Ambrosia, Artemicia, or Urticales;
(f) said fungus taxonomically belonging to Alternaria belongs taxonomically to Alternata;
(g) said fungus taxonomically belonging to Cladosporium belongs taxonomically to herbarium;
(h) said dust mite belonging taxonomically belonging to Dermatophagoides belongs taxonomically to Farinae or Pteronyssinus;
(i) said animal taxonomically belonging to Felis belongs taxonomically to Domesticus;
(j) said animal taxonomically belonging to Canis belongs taxonomically to familiaris;
(k) said animal taxonomically belonging to Equus belongs taxonomically to caballus;
(l) said insect taxonomically belonging to Blatella belongs taxonomically to Germanica;
(m) said insect taxonomically belonging to Periplaneta belongs taxonomically to americana;
(n) said insect taxonomically belonging to Hymenoptera belongs taxonomically to Vespidae, Apidae; and
(o) said weed taxonomically belonging to Urticales belongs taxonomically to Parietaria.
23. The recombinant allergen of claim 20, wherein the recombinant allergen is derived from Ves v 5.
24. The recombinant allergen of claim 23, wherein the allergen has one or more amino acid substitutions selected from the group consisting of:
i) Ala at position 72 of SEQ ID NO: 39; and
ii) Ala at position 96 of SEQ ID NO: 39.
25. The recombinant allergen of claim 20, wherein the recombinant allergen is derived from Bet v 1.
26. The recombinant allergen of claim 25, wherein the allergen has one or more amino acid substitutions selected from the group consisting of:
(i) Pro at position 10 of SEQ ID NO: 37;
(ii) Gly at position 25 of SEQ ID NO: 37;
(iii) Thr at position 28 of SEQ ID NO: 37, Gln at position 32 of SEQ ID NO: 37;
(iv) Ser at position 45 of SEQ ID NO: 37;
(v) Ser at position 47 of SEQ ID NO: 37;
(vi) Asn at position 55 of SEQ ID NO: 37;
(vii) Ala at position 77 of SEQ ID NO: 37;
(viii) Gly at position 108 of SEQ ID NO: 37; and
(ix) Thr at position 28 of SEQ ID NO: 37, Gln at position 32 of SEQ ID NO: 37, Ser at position 45 of SEQ ID NO: 37, and Gly at position 108 of SEQ ID NO: 37.
27. The recombinant allergen of claim 13, which is produced by a method comprising:
a) identifying amino acid residues in said naturally occurring allergen which are conserved with more than 70% identity in all of the known homologous proteins within the taxonomic order from which said naturally occurring allergen originates;
b) defining at least one patch of said conserved amino acid residues which patch (i) is connected over at least 400 2 of the surface of the three-dimensional structure of said naturally occurring allergen molecule; and (ii) comprises at least one B cell epitope, and wherein (iii) said conserved amino acids have a solvent accessibility of at least 20%; and
c) substituting at least one of said identified amino acid residues in said defined at least one patch with a non-conservative amino acid residue.
28. A pharmaceutical composition comprising a recombinant allergen prepared according to claim 1 and a pharmaceutically acceptable carrier.
29. A pharmaceutical composition comprising the recombinant allergen according to claim 13 and a pharmaceutically acceptable carrier.
30. A method of generating an immune response in a subject, which method comprises administering to the subject at least one recombinant allergen prepared according to claim 1 or a pharmaceutically acceptable composition comprising said at least one recombinant allergen.
31. A method of generating an immune response in a subject, which method comprises administering to the subject at least one recombinant allergen according to claim 13 or a pharmaceutically acceptable composition comprising said at least one recombinant allergen.
32. A method of vaccination or treatment of a subject, which method comprises administering to the subject at least one recombinant allergen prepared according to claim 1 or a pharmaceutically acceptable composition comprising said at least one recombinant allergen.
33. A method of vaccination or treatment of a subject, which method comprises administering to the subject at least one recombinant allergen according to claim 13 or a pharmaceutically acceptable composition comprising said at least one recombinant allergen.
34. A method of treatment, prevention or alleviation of allergic reactions in a subject, which method comprises administering to the subject at least one recombinant allergen prepared according to claim 1 or a pharmaceutically acceptable composition comprising said at least one recombinant allergen.
35. A method of treatment, prevention or alleviation of allergic reactions in a subject, which method comprises administering to the subject at least one recombinant allergen according to claim 13 or a pharmaceutically acceptable composition comprising said at least one recombinant allergen.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A light emitting diode (LED) driving apparatus which drives a plurality of LED arrays, the LED driving apparatus comprising:
a DC-DC converter which provides a driving voltage to the plurality of LED arrays;
a plurality of switching units, each of the switching units being connected in series to a corresponding LED array of the plurality of LED arrays and varying a size of a driving current to flow in the corresponding LED array of the plurality of LED arrays; and
a control unit which, in order for each of the switching units to operate within a predetermined headroom voltage range, calculates a driving current of each of the plurality of LED arrays and a duty cycle of the switching units corresponding to each of the plurality of LED arrays, and controls the plurality of switching units based on the calculated driving current and the calculated duty cycle,
wherein the control unit receives brightness information with respect to each of the plurality of LED arrays, calculates an average driving current of each of the plurality of LED arrays based on the received brightness information, calculates the driving current to be supplied to each of the plurality of LED arrays so that each of the corresponding plurality of switching units operate within the predetermined headroom voltage range, calculates a duty cycle of each of the plurality of switching units based on the calculated average driving current and the calculated driving current, and controls the plurality of switching units based on the calculated driving current and the calculated duty cycle.
2. The LED driving apparatus of claim 1, wherein
the control unit controls the DC-DC converter to provide the plurality of LED arrays with a sum of a forward voltage of an LED array having a largest forward voltage among the plurality of LED arrays and a lower voltage of the predetermined headroom voltage range as the driving voltage.
3. The LED driving apparatus of claim 1, wherein the control unit comprises:
a reference controller which calculates an average driving current of each of the plurality of LED arrays and generates an upper voltage value and a lower voltage value of the predetermined headroom voltage range;
an average current controller which calculates a driving current of each of the plurality of LED arrays by using the calculated average driving current, and further calculates a duty cycle of the switching units corresponding to each of the plurality of LED arrays by using the calculated average driving current, the driving current and the duty cycle being calculated in order for each of the plurality of switching units to operate between the upper voltage value and the lower voltage value; and
a plurality of current controllers, each of the current controllers controlling a corresponding switching unit of the plurality of switching units based on the calculated driving current and the calculated duty cycle.
4. The LED driving apparatus of claim 3, wherein
the reference controller receives brightness information with respect to each of the plurality of LED arrays, and calculates the average driving current of each of the plurality of LED arrays based on the received brightness information.
5. The LED driving apparatus of claim 3, wherein:
the average current controller controls the DC-DC converter to supply the calculated driving voltage to the plurality of LED arrays.
6. The LED driving apparatus of claim 5, wherein
the average current controller controls the DC-DC converter to supply a sum of a forward voltage of the LED array having a largest forward voltage among the plurality of LED arrays and a lower voltage of the predetermined headroom voltage range as the driving voltage with respect to the plurality of LED arrays.
7. The LED driving apparatus of claim 3, wherein:
the average current controller calculates the driving current of each of the plurality of LED arrays by using the calculated average driving current in order for each of the plurality of switching units to operate between the upper voltage value and the lower voltage value, and calculates the duty cycle of each of the plurality of switching units based on the average driving current calculated by the reference controller and the calculated driving current.
8. The LED driving apparatus of claim 7, wherein
in order for each of the plurality of switching units to operate between the upper voltage value and the lower voltage value, the average current controller calculates a driving current having a value which is increased compared to the average driving current calculated with respect to the other LED arrays except for the LED array having a largest forward voltage among the plurality of LED arrays.
9. The LED driving apparatus of claim 8, wherein
the average current controller calculates a duty cycle which is lower than a duty cycle of the LED array having the largest forward voltage with respect to the other LED arrays except for the LED array having the largest forward voltage among the plurality of LED arrays.
10. The LED driving apparatus of claim 3, wherein each of the plurality of switching units comprises:
a resistor comprising one end which is grounded; and
a switching component connected in series between the corresponding LED array and the resistor.
11. The LED driving apparatus of claim 10, wherein:
the switching component comprises a bipolar junction transistor (BJT) comprising a collector which is connected to one end of the corresponding LED array, a base connected to the corresponding current controller, and an emitter grounded through the resistor; and
each of the plurality of current controllers provides the base of the corresponding BJT with a value of voltage corresponding to the calculated driving current.
12. The LED driving apparatus of claim 10, wherein:
the switching component comprises a field effect transistor (FET) comprising a drain connected to one end of the corresponding LED array, a gate connected to the corresponding current controller, and a source grounded through the resistor, and
each of the plurality of current controllers provides the gate of the corresponding FET with a value of voltage corresponding to the calculated driving current.
13. The LED driving apparatus of claim 10, wherein:
each of the current controllers performs feedback control of a driving current to flow in the corresponding LED array by using a voltage value of the resistor.
14. The LED driving apparatus of claim 13, wherein
the control unit further comprises a plurality of comparators corresponding to the plurality of switching units, each of the comparators outputting a difference between the value of voltage corresponding to the driving current calculated in the average current controller and the voltage value of the resistor in the corresponding switching unit.
15. The LED driving apparatus of claim 14, wherein
each of the plurality of current controllers performs feedback control of a driving current to flow in the corresponding LED array based on an output of the corresponding comparator.
16. A light emitting diode (LED) driving method of an LED driving apparatus which comprises a plurality of LED arrays and a plurality of switching units configured to vary a size of a driving current to flow in each of the plurality of LED arrays, the LED driving method comprising:
receiving brightness information with respect to each of the plurality of LED arrays;
calculating an average driving current of each of the plurality of LED arrays based on the received brightness information;
calculating a driving current of each of the plurality of LED arrays in order for each of the plurality of switching units to operate within a predetermined headroom voltage range;
calculating a duty cycle of each of the plurality of switching units based on the calculated average driving current and the calculated driving current; and
controlling the plurality of switching units based on the calculated driving current and the calculated duty cycle.
17. The LED driving method of claim 16, wherein
the calculating of the driving current comprises calculating the driving current to have a value which is increased compared to the average driving current calculated with respect to the other LED arrays except for an LED array having a largest forward voltage among the plurality of LED arrays in order for each of the plurality of switching units to operate within the predetermined headroom voltage range.
18. The LED driving method of claim 17, wherein
the calculating of the duty cycle comprises calculating the duty cycle to be lower than a duty cycle of the LED array having the largest forward voltage with respect to the other LED arrays except for the LED array having the largest forward voltage among the plurality of LED arrays.
19. A non-transitory computer-readable recording medium comprising a program for executing a light emitting diode (LED) driving method in an LED driving apparatus which comprises a plurality of LED arrays and a plurality of switching units configured to vary a size of a driving current to flow in each of the plurality of LED arrays, the non-transitory computer-readable recording medium causing a computer to execute the LED driving method comprising:
receiving brightness information with respect to each of the plurality of LED arrays;
calculating an average driving current of each of the plurality of LED arrays based on the received brightness information;
calculating a driving current of each of the plurality of LED arrays in order for each of the plurality of switching units to operate within a predetermined headroom voltage range;
calculating a duty cycle of each of the plurality of switching units based on the calculated average driving current and the calculated driving current; and
controlling the plurality of switching units based on the calculated driving current and the calculated duty cycle.