Anatomy of the antibody molecule

Padlan, Eduardo A.

doi:10.1016/0161-5890(94)90001-9

Cited by 850 publications

(628 citation statements)

References 142 publications

Supporting

Mentioning

591

Contrasting

Unclassified

Order By: Relevance

“…That more stringent selection for proteolytic stability results in the selection of clones from a single CDR group suggests that CDRs are the primary site of proteolysis. This is in accordance with the general view that CDRs form flexible surface-exposed loops (Padlan 1994;Harmsen et al 2000), which is a structure that is more susceptible to proteolysis (North 1989). However, the newly selected clones still varied considerably in proteolytic stability.…”

Section: Discussionsupporting

confidence: 90%

“…A scattered pattern of amino acid substitutions is commonly found in recombinant antibodies selected for improved affinity or physicochemical stability (Daugherty et al 2000;Jermutus et al 2001;Zahnd et al 2004). It has been reported that amino acid substitutions outside the CDRs that result in increased affinity can stabilize CDR loops in a conformation that is more suitable for antigen binding (Foote and Winter 1992;Padlan 1994;Wedemayer et al 1997). Possibly, the higher proteolytic stability of the newly selected VHHs is similarly caused by reduced flexibility of CDR loops.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy

Harmsen

Solt

Bemmel³

et al. 2006

Appl Microbiol Biotechnol

107

View full text Add to dashboard Cite

We previously demonstrated that oral application of the recombinant single-domain antibody fragment (VHH) clone K609, directed against Escherichia coli F4 fimbriae, reduced E. coli-induced diarrhoea in piglets, but only at high VHH doses. We have now shown that a large portion of the orally applied K609 VHH is proteolytically degraded in the stomach. Stringent selection for proteolytic stability identified seven VHHs with 7-to 138-fold increased stability after in vitro incubation in gastric fluid. By DNA shuffling we obtained four clones with a further 1.5-to 3-fold increased in vitro stability. These VHHs differed by at most ten amino acid residues from each other and K609 that were scattered over the VHH sequence and did not overlap with predicted protease cleavage sites. The most stable clone, K922, retained 41% activity after incubation in gastric fluid and 90% in jejunal fluid. Oral application of K922 to piglets confirmed its improved proteolytic stability. In addition, K922 bound to F4 fimbriae with higher affinity and inhibited fimbrial adhesion at lower VHH concentrations. K922 is thus a promising candidate for prevention of piglet diarrhoea. Furthermore, our findings could guide selection and improvement by genetic engineering of other recombinant antibody fragments for oral use.

show abstract

Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy

Harmsen

Solt

Bemmel³

et al. 2006

Appl Microbiol Biotechnol

107

View full text Add to dashboard Cite

show abstract

“…In one, the four different isotypes provide different combinations of V L CDR1 and CDR2 length which influence the binding site topology, probably by specific support of V H CDR3; those V L with longer CDR2 such as r associated with V H having short CDR3 [36]. This hypothesis would predict that V H genes expressed at different stages of development or in different tissues would show an L chain preference.…”

Section: Discussion Natural History Of Four Ancient Igl Cladesmentioning

confidence: 99%

Four primordial immunoglobulin light chain isotypes, including λ and κ, identified in the most primitive living jawed vertebrates

Criscitiello

Flajnik

2007

Eur J Immunol

View full text Add to dashboard Cite

The discovery of a fourth immunoglobulin (Ig) light (L) chain isotype in sharks has revealed the origins and natural history of all vertebrate L chains. Phylogenetic comparisons have established orthology between this new shark L chain and the unique Xenopus L chain isotype r. More importantly, inclusion of this new L chain family in phylogenetic analyses showed that all vertebrate L chains can be categorized into four ancestral clans originating prior to the emergence of cartilaginous fish: one restricted to elasmobranchs (r-cart/type I), one found in all cold-blooded vertebrates (r/teleost type 2/elasmobranch type IV), one in all groups except bony fish (k/elasmobranch type II), and one in all groups except birds (j/elasmobranch type III/teleost type 1 and 3). All four of these primordial L chain isotypes (r, r-cart, k and j) have maintained separate V region identities since their emergence at least 450 million years ago, suggestive of an ancient physiological distinction of the L chains. We suggest that, based upon unique, discrete sizes of complementarity determining regions 1 and 2 and other features of the V region sequences, the different L chain isotypes arose to provide different functional conformations in the Ig binding site when they pair with heavy chains.

show abstract

“…Imprecision in gene segment joining and variation in the extent of N nucleotide insertion create a CDR-H3 repertoire that ranges from unmodified and intact germ-lineencoded sequence to sequences where extensive nibbling and N addition have obscured the identity of the D H progenitor. The broad range of diversity available to CDR-H3 has functional consequences, because its location at the center of the antigen binding site, as classically defined, permits this interval to often play a significant role in antigen recognition and binding [3][4][5].…”

Section: Introductionmentioning

confidence: 99%

Categorical selection of the antibody repertoire in splenic B cells

et al. 2007

View full text Add to dashboard Cite

In the bone marrow, the passage of developing B cells through critical checkpoints of differentiation is associated with a reduction of specific categories of CDR3 of the Ig heavy chain (CDR-H3), particularly those with excessive hydrophobic or charged amino acids and those with a length of eight or fewer residues. To gain insight into the role of CDR-H3 content in the development of B cells in the spleen, we compared the sequences of V H 7183DJCl transcripts from sorted transitional T1, marginal zone, and follicular B cell subsets to those expressed by immature IgM + IgD -and mature IgM lo IgD hi B cells in the bone marrow. Although differences in V H utilization were noted, the T1 CDR-H3 repertoire showed extensive similarity to that of immature bone marrow B cells, and the follicular CDR-H3 repertoire most resembled that of mature bone marrow B cells. Unlike the splenic follicular and bone marrow mature B cell CDR-H3 repertoires, the marginal zone B cell CDR-H3 repertoire retained both short and highly charged amino acid motifs, including those with two arginines. Our findings suggest that antigen binding sites containing specific categories of CDR-H3 sequence content may inhibit, permit, or even facilitate passage of the host B cell through critical checkpoints in peripheral as well as central development.

show abstract

Anatomy of the antibody molecule

Abstract: The structures of the various regions of an antibody molecule are analysed and correlated with biological function. The structural features which relate to potential applications are detailed.

Cited by 850 publications

References 142 publications

Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy

Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy

Four primordial immunoglobulin light chain isotypes, including λ and κ, identified in the most primitive living jawed vertebrates

Categorical selection of the antibody repertoire in splenic B cells

Contact Info

Product

Resources

About