The assignment of amino acids to each region or domain is in
accordance with the definitions of Kabat Sequences of Proteins of Immunological
Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)) or in
10 accordance with the definitions of Chothia et al. Conformations of immunoglobulin hypervariable regions (Nature 1989; 342(6252)877-83). The term "antibody"
encompasses murine, ฮิวเมไนซ์, ฮิวเมน and ไคเมริก antibodies, and an antibody
that is a multimeric form of antibodies, such as ไดเมอร์, trimers, or higher-order
multimers of monomeric antibodies. Antibody also encompasses โมโนสเปซิฟืก,
15 ไบสเปซิฟืก or มัลติสเปซิฟืก antibodies, and any other modified configuration of the
immunoglobulin molecule that ประกอบรวมด้วย an antigen recognition site of the
required specificity. It also encompasses an antibody that is linked or attached to a
non-antibody moiety. Further, the term "antibody" is not limited by any particular
method of producing the antibody. For example, it รวมถึง โมโนโคลนอล antibodies, 20 รีคอมบิแนนต์ antibodies and โพลีโคลนอล antibodies.
The term "antibody derivative" or "derivative" of an antibody refers to a
molecule that is capable of binding to the same antigen (i.e., ฮิวเมน CD134) that
the antibody binds to and ประกอบรวมด้วย an amino acid sequence of the antibody linked
to an additional molecular entity. The amino acid sequence of the antibody that is 25 contained in the antibody derivative may be the full-length antibody, or may be
any portion or portions of a full-length antibody. The additional molecular entity
may be a biological or chemical molecule. Examples of additional molecular entities
รวมถึง chemical groups, peptides, proteins (such as enzymes, antibodies), amino
acids, and chemical compounds. The additional molecular entity may be for use as 30 a detection agent, marker label, therapeutic or pharmaceutical agent. The amino
acid sequence of an antibody may be attached or linked to the additional entity by
non-covalent association, chemical coupling, genetic fusion, or otherwise. The term
"antibody derivative" also encompasses ไคเมริก antibodies, ฮิวเมไนซ์ antibodies,
and molecules that are derived from การดัดแปลง of the amino acid sequences of a CD134 antibody, such as conservation amino acid substitutions, insertions and additions.
The term "antigen-binding fragment" of an antibody refers to one or more
5 portions of a full-length antibody that retain the ability to bind to the same antigen
(i.e., ฮิวเมน CD134) that the antibody binds to. The term "antigen-binding
fragment" also encompasses a portion of an antibody that is part of a larger
molecule formed by non-covalent or covalent association or of the antibody portion
with one or more additional molecular entities. Examples of additional molecular 10 entities รวมถึง amino acids, peptides, or proteins, such as the สเตร็ปทาวิดิน core
region, which may be used to make a tetrameric scFv molecule (Kipriyanov et al.
Hum Antibodies Hybridomas 1995; 6(3): 93-101). An exemplary antigen-binding
fragment is a VH and/or a VL of an antibody.
The term "ไคเมริก antibody" refers to an antibody that ประกอบรวมด้วย amino 15 acid sequences derived from two different species such as ฮิวเมน and mouse,
typically a combination of mouse variable (from heavy and light chains) regions
and ฮิวเมน constant (heavy and light chains) regions .
The term "เอพิโทป" refers to the part of an antigen that is capable of specific
binding to an antibody, or T-เซลล์ รีเซปเตอร์ or otherwise interacting with a molecule. 20 "เอพิโทป" is also referred to in the art as the "antigenic determinant". An เอพิโทป
generally consists of chemically active surface groupings of molecules such as
amino acids or carbohydrate or sugar side chains. An เอพิโทป may be "linear" or
"non-linear/conformational". Once a desired เอพิโทป is determined (e.g., by เอพิโทป
mapping), antibodies to that เอพิโทป can be generated. The generation and
25 characterization of antibodies may also provide information about desirable
เอพิโทป. From this information, it is then possible to screen antibodies for those
which bind to the same เอพิโทป e.g. by conducting cross-competition studies to find
antibodies that competitively bind with one another, i.e., the antibodies compete for
binding to the antigen.
30 The term "host cell" refers to a cell into which an expression vector has been
introduced. The term encompasses not only the particular subject cell but also the progeny of such a cell. Because certain การดัดแปลง may occur in successive
generations due to either environmental influences or mutation, such progeny may
not be identical to the parent cell, but are still รวมถึงd within the scope of the term "host cell."
The term "ฮิวเมน antibody" refers to an antibody consisting of amino acid
sequences of ฮิวเมน immunoglobulin sequences only. A ฮิวเมน antibody may
5 contain murine carbohydrate chains if produced in a mouse, in a mouse cell or in a
hybridoma derived from a mouse cell. ฮิวเมน antibodies may be prepared in a
variety of ways known in the art.
The term "ฮิวเมไนซ์ antibody" refers to an antibody that contains some or
all of the CDRs from a non-ฮิวเมน animal antibody while the framework and
10 constant regions of the antibody contain amino acid residues derived from ฮิวเมน
antibody sequences. ฮิวเมไนซ์ antibodies are typically produced by grafting
CDRs from a mouse antibody into ฮิวเมน framework sequences followed by back
substitution of certain ฮิวเมน framework residues for the corresponding mouse
residues from the source antibody. The term "ฮิวเมไนซ์ antibody" also refers to 15 an antibody of non-ฮิวเมน origin in which, typically in one or more variable
regions, one or more เอพิโทป have been removed, that have a high propensity of
constituting a ฮิวเมน T-เซลล์ and/or B-cell เอพิโทป, for purposes of reducing
immunogenicity. The amino acid sequence of the เอพิโทป can be removed in full or
in part. However, typically the amino acid sequence is altered by substituting one 20 or more of the amino acids constituting the เอพิโทป for one or more other amino
acids, thereby changing the amino acid sequence into a sequence that does not
constitute a ฮิวเมน T-เซลล์ and/or B-cell เอพิโทป. The amino acids are substituted by
amino acids that are present at the corresponding position(s) in a corresponding
ฮิวเมน variable heavy or variable light chain as the case may be.
25 The term "mammal" refers to any animal species of the Mammalian class.
Examples of mammals รวมถึง: ฮิวเมนs; laboratory animals such as rats, mice, simians and guinea pigs; domestic animals such as rabbits, cattle, sheep, goats, cats, dogs, horses, and pigs and the like.
The term "isolated nucleic acid" refers to a nucleic acid molecule of cDNA, or 30 synthetic origin, or a combination ของมัน, which is separated from other nucleic
acid molecules present in the natural source of the nucleic acid.
The term "Ka" refers to the equilibrium dissociation constant of a particular antibody-antigen interaction and is used to describe the binding affinity between a
16
ลิแกนด์ (such as an antibody) and a protein (such as CD134). The smaller the
equilibrium dissociation constant, the more tightly bound the ลิแกนด์ is, or the
higher the affinity between ลิแกนด์ and protein. A Ka can be measured by surface
plasmon resonance, for example using the BIACORE 1 or the Octet system. The 5 term "แอนติ-CD134 antibody" refers to an antibody, as defined herein, capable of
binding to the ฮิวเมน CD134.
The terms "OX40 รีเซปเตอร์", "CD134 รีเซปเตอร์" and "CD134" are used
interchangeably in the present application, and รวมถึง the ฮิวเมน CD134, as well
as variants, isoforms, and species homologues ของมัน. Accordingly, ฮิวเมน CD134 10 โมเลกุลยึดเกาะs disclosed herein may, in certain cases, also bind to the CD134
from species other than ฮิวเมน. For example, the โมเลกุลยึดเกาะs of the
การประดิษฐ์ may have cross-reactivity to other related antigens, for example to the
CD134 from other species, such as ฮิวเมน or monkey, for example Macaca
fascicularis (cynomolgus, cyno) or Pan troglodytes (chimpanzee, chimp). In other 15 cases, the โมเลกุลยึดเกาะs may be completely specific for the ฮิวเมน CD134 and
may not exhibit species or other types of cross-reactivity. For example, they will
not bind to the mouse or rat CD134.
The term "specifically bind to the ฮิวเมน CD134" means that the Ka of a
โมเลกุลยึดเกาะ for binding to ฮิวเมน CD134, is less than about 10 fold, 50 fold 20 or100 fold the Ka for its binding to, e.g., the ฮิวเมน CD40, as determined using an
assay described herein or known to one of skill in the art (e.g. a BlAcore assay).
The determination that a particular agent binds specifically to the OX40
รีเซปเตอร์ may alternatively readily be made by using or adapting routine
procedures. One suitable in vitro assay makes use of the Western blotting
25 procedure (described in many standard texts, ที่รวมถึง "Antibodies, A Laboratory
Manual" by Harlow and Lane).