Functional Assurance and Evidence-based Results

Potency, or bio-functional status, of an active principle is dependent on molecular separation (isolation) and structural stability of a bioactive compound. Protein engineering to map the three-dimensional structures of bioactive molecules is a specialized field. bioQuad® Life Sciences is a leading world expert in protein engineering. The first activation of any protein structure outside an in vivo system was first reported by Dr. Naidu and his team in the year 2000 with ‘Activated Lactoferrin’ – a research finding that made global headlines. The purity (certificate of analysis) and homogeneity of a biochemical does not necessarily assure the potency of such molecule. Bioactive molecules ‘unfold and die’; therefore, their stability data and expiry date (best before use) is disclosed for nutritional ingredients. Our laboratory routinely performs biological testing of active ingredients and their finished products to ensure their functional performance. ‘Quality Assurance’ is an important GMP protocol, but we also believe that ‘Functional Assurance’ (which is not a standard norm practiced in the health supplement industry) is much more critical to ensure the biological performance (potency) of the product.

Lactoferrin Treated for Contaminant Reduction (LF-TCR)

patented 3 tier filtration process of milk lactoferrin

Lactoferrin is a milk protein credited with an impressive list of health benefits. Separation of lactoferrin from other ingredients of milk requires several complex steps of protein engineering. Despite high purity, proteins isolated from such processes may harbor microbial and endotoxin contaminants that could compromise lactoferrin functionality. A novel treatment for contaminant reduction (TCR) to enhance the protein quality during commercial-scale lactoferrin production has been developed. Lactoferrin / LF-(TCR), based on this cutting-edge protein technology, contains ultra-clean lactoferrin that could offer the highest standards of microbiological quality and functional assurance for nutraceutical applications.

Intestinal Maintenance

Background - The human intestinal tract (consisting of small and large intestines) is a long (30') semi-permeable biological tube that separates the true inside of the body from the outside. Its major functions include to i) absorb digested food (nutrients, energy, water), ii) excrete waste material (feces) and iii) keep out pathogens (serve as a defense barrier). The lining of the intestine is covered with microvilli (tiny projections) that exponentially increase the surface area of the intestinal mucosa. The integrity and density of microvilli is vital for optimal absorption of nutrients.

human body showing intestinal lining

Study Objective - To preserve the integrity and density of the intestinal microvilli, which is vital for efficient nutrient absorption.

intestinal cell lining preserved by lactoferrin TCR

Study Results - In this CaCO2 cell culture experiment, intestinal epithelia cells were grown as monolayers in vitro. The monolayers were visualized under phase-contrast microscopy for integrity and morphology. Normal viable cell monolayer appear confluent, compact, round and bright. We observed the cell morphology over 24 hours. There were four cell culture groups - CONTROL (cells without any treatment); and cells treated with i) whey LF, ii) milk LF and iii) milk LF-(TCR). After 24 hours, as shown - the CONTROLS showed cell death (apoptosis with dark brown shade). Monolayers treated with whey and milk LF actually showed both apoptosis and cytotoxicity resulting from protein exposure (dark murky color). In contrast, monolayers treated with LF-(TCR) showed confluent cell growth with healthy structural morphology (bright color with no discoloration).

Reference: Naidu AS (2005) Ultra-cleansing of lactoferrin: Nutraceutical implications. Eur J Nutraceut Funct Foods 16 (March/April):7-13.

Host Defense

Background - The human host defense operates with two distinct pathways: i) the first-line (or non-specific) defense against invading organisms; and ii) the adaptive (or specific) immune-type, which acts as a second-line of defense to provide protection against re-exposure to the same pathogen. The interplay between these two arms of immune system establishes an effective host defense.

immune cells engulfing microbes

Study Objective - To inhibit the growth of pathogenic bacteria, thereby minimizing complications associated with infection.

bacteria inhibited by lactoferrin TCR

Study Results - In this experiment, we have compared the ability of different LF preparations to inhibit (via bacteriostasis) the growth of pathogenic E.coli. Green-fluorescent protein (GFP)-labelled E. coli were exposed to 1% LF containing broth solutions for 24 h. Broth without any LF served as positive growth CONTROL. The intensity of GFP (measured by florescent microscopic imaging) has reflected the extent of bacterial growth-multiplication. Thus, higher the bacterial growth, brighter the green color as visualized in the CONTROL broth (without LF). Normal LF showed a slight decrease in the green fluorescence. In contrast, the LF-(TCR) has completely inhibited the growth-multiplication of the pathogenic E.coli - resulting in no enhancement of the green fluorescence. 

Reference: Naidu AS (2005) Ultra-cleansing of lactoferrin: Nutraceutical implications. Eur J Nutraceut Funct Foods 16 (March/April):7-13.

Ribonuclease-Enriched Lactoferrin (RELF)

Bone Turnover

Background - The bone is a dynamic tissue that undergoes a continuous remodeling process through repeated cycles of destruction and rebuilding. In healthy adults, the amount of new bone formation approximately balances the amount of bone resorption. As age increases, and due to factors such as diet, lifestyle, medications, etc., the balance shifts to favor bone resorption. A reduction in angiogenesis could also contribute to this imbalance. Maintaining healthy bone turnover is essential to reinforce skeletal structure and optimize its function.

hip bone with bone matrix background

Study Objective - Examine the effect of RNase-enriched lactoferrin (R-ELF) supplement on bone turnover (i.e. bone formation and bone resorption).

bone resorption decreased and bone formation increased by ribonuclease-enriched lactoferrin

Study Results - A total of 35 healthy, post-menopausal women, aged 45 to 60 years, were randomized into placebo or R-ELF supplement group. Over a six-month period, the bone status was monitored by assessing bone resorption markers, serum N-telopeptides (sNTx) and urine deoxypyridinoline (uDpd) cross-links; and serum bone formation markers, bone-specific alkaline phosphatase (SBAP) and osteocalcin (sOC). R-ELF supplmentation demonstrated a decrease in uDpD levels by 14% (19% increase in placebo) and sNTx maintained at 25% of the baseline (41% for placebo), while serum BAP and OC levels showed a 45% and 16% elevation (25% and 5% for placebo). R-ELF supplementation showed a statistically signifcant reduction in bone resorption and increase in osteoblastic bone formation, to restore the balance of bone turnover within a short period.

Reference: Bharadwaj S, Tezus Naidu AG, Betagiri GV, Prasadarao NV, Naidu AS (2009) Milk ribonuclease-enriched lactoferrin induces positive effects on bone turnover markers in postmenopausal women. Osteoporos Int 20(9):1603-1611.

Joint Inflammation

Background - Bone and joints work together to enable movement, similar to a joist and lever system. For optimal function, the joints have to preserve structure, maintain lubrication (i.e. synovial fluid), remove debris (normal by-product of wear-and-tear) and minimize inflammation. Inflammation is the primary defense mechanism fo the body characterized by a 'high alert' state of the immune system, triggered by the release of several pro-inflammatory cytokines. Excessive inflammation can be injurious to health; in the case of joints, it can damage tissues, induce pain, and restrict movement.

human body showing joint inflammation process

Study Objective - Evaluate the effect of RNase-enriched lactoferrin (R-ELF) supplement on joint inflammation (by measuring circulating cytokine levels).

pro-inflammation decreased and anti-inflammation increased by ribonuclease-enriched lactoferrin

Study Results - A six-month, randomized clinical study was conducted on 35 healthy, post-menopausal women, aged 45 to 60 years. They were randomized into placebo or R-ELF supplement group; the R-ELF group was supplemented with R-ELF (2x 125 mg/day) and calcium (100% RDA), while the placebo group received only the calcium supplement. Serum levels of receptor activator for NF-κB ligand (RANKL), C-reactive protein (CRP) and various pro- and anti-inflammatory cytokines were determined by ELISA. Pro-inflammatory cytokines IL-6 and TNF-α decreased significantly (-44% and -10% respectively) while anti-inflammatory IL-10 increased (140%) with R-ELF supplementation. RANKL and CRP were modestly reduced (-50%) relative to the placebo, while RANKL elevated initially. R-ELF supplementation showed beneficial effects towards improvement of inflammatory status.

Reference: Bharadwaj S, Tezus Naidu AG, Betagiri GV, Prasadarao NV, Naidu AS (2010) Inflammatory responses improve with milk ribonuclease-enriched lactoferrin supplementation in postmenopausal women. J Inflammation Res 59(11):971-978.