| Title |
Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy
|
|---|---|
| Published in |
Applied Microbiology and Biotechnology, February 2006
|
| DOI | 10.1007/s00253-005-0300-7 |
| Pubmed ID | |
| Authors |
M. M. Harmsen, C. B. van Solt, A. M. van Zijderveld-van Bemmel, T. A. Niewold, F. G. van Zijderveld |
| Abstract |
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. |
Mendeley readers
The data shown below were compiled from readership statistics for 150 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 2 | 1% |
| Brazil | 1 | <1% |
| Portugal | 1 | <1% |
| Belgium | 1 | <1% |
| United Kingdom | 1 | <1% |
| Unknown | 144 | 96% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 41 | 27% |
| Student > Ph. D. Student | 32 | 21% |
| Student > Master | 18 | 12% |
| Student > Bachelor | 17 | 11% |
| Other | 5 | 3% |
| Other | 14 | 9% |
| Unknown | 23 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 49 | 33% |
| Biochemistry, Genetics and Molecular Biology | 36 | 24% |
| Immunology and Microbiology | 13 | 9% |
| Medicine and Dentistry | 6 | 4% |
| Pharmacology, Toxicology and Pharmaceutical Science | 5 | 3% |
| Other | 14 | 9% |
| Unknown | 27 | 18% |
Attention Score in Context
This research output has an Altmetric Attention Score of 32. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 19 March 2024.
All research outputs
#1,156,496
of 24,119,703 outputs
Outputs from Applied Microbiology and Biotechnology
#50
of 8,034 outputs
Outputs of similar age
#2,906
of 159,944 outputs
Outputs of similar age from Applied Microbiology and Biotechnology
#2
of 60 outputs
Altmetric has tracked 24,119,703 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 95th percentile: it's in the top 5% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 8,034 research outputs from this source. They receive a mean Attention Score of 4.3. This one has done particularly well, scoring higher than 99% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 159,944 tracked outputs that were published within six weeks on either side of this one in any source. This one has done particularly well, scoring higher than 98% of its contemporaries.
We're also able to compare this research output to 60 others from the same source and published within six weeks on either side of this one. This one has done particularly well, scoring higher than 96% of its contemporaries.