Multivalent Anchoring and Oriented Display of Single-Domain Antibodies on Cellulose

Antibody engineering has allowed for the rapid generation of binding agents against virtually any antigen of interest, predominantly for therapeutic applications. Considerably less attention has been given to the development of diagnostic reagents and biosensors using engineered antibodies. Recently, we produced a novel pentavalent bispecific antibody (i.e., decabody) by pentamerizing two single-domain antibodies (sdAbs) through the verotoxin B subunit (VTB) and found both fusion partners to be functional. Using a similar approach, we have engineered a bispecific pentameric fusion protein consisting of five sdAbs and five cellulose-binding modules (CBMs) linked via VTB. To find an optimal design format, we constructed six bispecific pentamers consisting of three different CBMs, fused to the Staphylococcus aureus-specific human sdAb HVHP428, in both orientations. One bispecific pentamer, containing an N-terminal CBM9 and C-terminal HVHP428, was soluble, non-aggregating, and did not degrade upon storage at 4 °C for over six months. This molecule was dually functional as it bound to cellulose-based filters as well as S. aureus cells. When impregnated in cellulose filters, the bispecific pentamer recognized S. aureus cells in a flow-through detection assay. The ability of pentamerized CBMs to bind cellulose may form the basis of an immobilization platform for multivalent display of high-avidity binding reagents on cellulosic filters for sensing of pathogens, biomarkers and environmental pollutants

Efficacy of a voluntary BVDV control programme: Experiences from the Netherlands

The outcomes of a voluntary bovine viral diarrhoea virus (BVDV) control programme that has been in place in the Netherlands since 1997 were analysed. This ‘BVDV-free’ programme was studied in dairy herds in the period 1 August 2007 to 1 August 2013. The programme was based on a test and cull approach at the herd level, after which the BVDV status was monitored by testing young stock for antibodies against BVDV or by antigen testing of newborn calves. One of the challenges of the programme was that, without any legislation or subsidies, farmers had to be motivated to pay all costs involved, with eradication of BVDV from their farm as the only incentive. During the study period, the percentage of dairy farms with a ‘BVDV-free’ status in the Netherlands increased from 13% to 24%, while the prevalence of active BVDV infections in Dutch dairy herds decreased. This may be related to the increasing number of participants in the ‘BVDV-free’ programme

Efficacy of a voluntary BVDV control programme: Experiences from the Netherlands

The outcomes of a voluntary bovine viral diarrhoea virus (BVDV) control programme that has been in place in the Netherlands since 1997 were analysed. This ‘BVDV-free’ programme was studied in dairy herds in the period 1 August 2007 to 1 August 2013. The programme was based on a test and cull approach at the herd level, after which the BVDV status was monitored by testing young stock for antibodies against BVDV or by antigen testing of newborn calves. One of the challenges of the programme was that, without any legislation or subsidies, farmers had to be motivated to pay all costs involved, with eradication of BVDV from their farm as the only incentive. During the study period, the percentage of dairy farms with a ‘BVDV-free’ status in the Netherlands increased from 13% to 24%, while the prevalence of active BVDV infections in Dutch dairy herds decreased. This may be related to the increasing number of participants in the ‘BVDV-free’ programme

The effect of bovine viral diarrhea virus introduction on milk production of Dutch dairy herds

Dairy cows are negatively affected by the introduction of bovine viral diarrhea virus (BVDV), and consequently, produce less milk. Existing literature on potential milk production losses is based on relatively outdated data and hardly evaluates milk production loss in relation to a new BVDV infection in a surveillance system. This study determined the annual and quarterly loss in milk production of BVDV introduction in 3,126 dairy herds participating in the Dutch BVDV-free program between 2007 and 2017. Among these herds, 640 were “breakdown-herds” that obtained and subsequently lost their BVDV-free status during the study period, and 2,486 herds obtained and retained their BVDV-free status during the study period. Milk yields before and after BVDV introduction were compared through annual and quarterly linear mixed models. The fixed variables for both models included herd type (breakdown-herd or free-herd), bovine viral diarrhea status (on an annual and quarterly basis), year, season, and a random herd effect. The dependent variable was the average daily milk yield on the test day. To define the possible BVDV-introduction dates, 4 scenarios were developed. In the default scenario, the date of breakdown (i.e., loss of the BVDV-free status) was assumed as the BVDV-introduction date. For the other 3 scenarios, the BVDV-introduction dates were set at 4, 6, and 9 mo before the date of breakdown, based on the estimated birth date of a persistently infected calf. In the default scenario, the loss in milk yield due to BVDV introduction occurred mainly in the first year after breakdown, with a reduction in yield of 0.08 kg/cow per day compared with the last year before breakdown. For the other 3 scenarios, the greatest yield reduction occurred in the second year after BVDV introduction, with a loss of 0.09, 0.09, and 0.1 kg/cow per day, respectively. For the first 4 quarters after BVDV introduction in the default scenario, milk yield loss was 0.14, 0.09, 0.02, and 0.08 kg/cow per day, respectively. These quarterly results indicated that milk yield loss was greatest in the first quarter after BVDV introduction. Overall, BVDV introduction had a negative, but on average a relatively small, effect on milk yield for herds participating in the BVDV-free program. This study will enable dairy farmers and policymakers to have a clearer understanding of the quantitative milk production effect of BVDV on dairy farms in a control program

The effect of bovine viral diarrhea virus introduction on milk production of Dutch dairy herds

Dairy cows are negatively affected by the introduction of bovine viral diarrhea virus (BVDV), and consequently, produce less milk. Existing literature on potential milk production losses is based on relatively outdated data and hardly evaluates milk production loss in relation to a new BVDV infection in a surveillance system. This study determined the annual and quarterly loss in milk production of BVDV introduction in 3,126 dairy herds participating in the Dutch BVDV-free program between 2007 and 2017. Among these herds, 640 were “breakdown-herds” that obtained and subsequently lost their BVDV-free status during the study period, and 2,486 herds obtained and retained their BVDV-free status during the study period. Milk yields before and after BVDV introduction were compared through annual and quarterly linear mixed models. The fixed variables for both models included herd type (breakdown-herd or free-herd), bovine viral diarrhea status (on an annual and quarterly basis), year, season, and a random herd effect. The dependent variable was the average daily milk yield on the test day. To define the possible BVDV-introduction dates, 4 scenarios were developed. In the default scenario, the date of breakdown (i.e., loss of the BVDV-free status) was assumed as the BVDV-introduction date. For the other 3 scenarios, the BVDV-introduction dates were set at 4, 6, and 9 mo before the date of breakdown, based on the estimated birth date of a persistently infected calf. In the default scenario, the loss in milk yield due to BVDV introduction occurred mainly in the first year after breakdown, with a reduction in yield of 0.08 kg/cow per day compared with the last year before breakdown. For the other 3 scenarios, the greatest yield reduction occurred in the second year after BVDV introduction, with a loss of 0.09, 0.09, and 0.1 kg/cow per day, respectively. For the first 4 quarters after BVDV introduction in the default scenario, milk yield loss was 0.14, 0.09, 0.02, and 0.08 kg/cow per day, respectively. These quarterly results indicated that milk yield loss was greatest in the first quarter after BVDV introduction. Overall, BVDV introduction had a negative, but on average a relatively small, effect on milk yield for herds participating in the BVDV-free program. This study will enable dairy farmers and policymakers to have a clearer understanding of the quantitative milk production effect of BVDV on dairy farms in a control program